Processes for preparing JAK inhibitors and related intermediate compounds

ABSTRACT

The present invention is related to processes for preparing chiral substituted pyrazolyl pyrrolo[2,3-d]pyrimidines of Formula III, and related synthetic intermediate compounds. The chiral substituted pyrazolyl pyrrolo[2,3-d]pyrimidines are useful as inhibitors of the Janus Kinase family of protein tyrosine kinases (JAKs) for treatment of inflammatory diseases, myeloproliferative disorders, and other diseases.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.12/687,623, filed Jan. 14, 2010, which claims the benefit of U.S. Ser.No. 61/144,991, filed Jan. 15, 2009, the disclosures of which areincorporated herein by reference in its entirety.

FIELD OF THE INVENTION

The present invention is related to processes for preparing chiralsubstituted pyrazolyl pyrrolo[2,3-d]pyrimidines and related syntheticintermediate compounds. The chiral substituted pyrazolylpyrrolo[2,3-d]pyrimidines are useful as inhibitors of the Janus Kinasefamily of protein tyrosine kinases (JAKs) for treatment of inflammatorydiseases, myeloproliferative disorders, and other diseases.

BACKGROUND

Protein kinases (PKs) are a group of enzymes that regulate diverse,important biological processes including cell growth, survival anddifferentiation, organ formation and morphogenesis, neovascularization,tissue repair and regeneration, among others. Protein kinases exerttheir physiological functions through catalyzing the phosphorylation ofproteins (or substrates) and thereby modulating the cellular activitiesof the substrates in various biological contexts. In addition to thefunctions in normal tissues/organs, many protein kinases also play morespecialized roles in a host of human diseases including cancer. A subsetof protein kinases (also referred to as oncogenic protein kinases), whendysregulated, can cause tumor formation and growth, and furthercontribute to tumor maintenance and progression (Blume-Jensen P et al,Nature 2001, 411(6835):355-365). Thus far, oncogenic protein kinasesrepresent one of the largest and most attractive groups of proteintargets for cancer intervention and drug development.

Protein kinases can be categorized as receptor type and non-receptortype. Receptor tyrosine kinases (RTKs) have an extracellular portion, atransmembrane domain, and an intracellular portion, while non-receptortyrosine kinases are entirely intracellular. The Janus kinase family ofprotein tyrosine kinases (JAKs) belong to the non-receptor type oftyrosine kinases and include family members: JAK1 (also known as Januskinase-1), JAK2 (also known as Janus kinase-2), JAK3 (also known asJanus kinase, leukocyte; JAKL; L-JAK and Janus kinase-3) and TYK2 (alsoknown as protein-tyrosine kinase 2).

The pathway involving JAKs and Signal Transducers and Activators ofTranscription (STATs) is engaged in the signaling of a wide range ofcytokines Cytokines are low-molecular weight polypeptides orglycoproteins that stimulate biological responses in virtually all celltypes. Generally, cytokine receptors do not have intrinsic tyrosinekinase activity, and thus require receptor-associated kinases topropagate a phosphorylation cascade. JAKs fulfill this function.Cytokines bind to their receptors, causing receptor dimerization, andthis enables JAKs to phosphorylate each other as well as specifictyrosine motifs within the cytokine receptors. STATs that recognizethese phosphotyrosine motifs are recruited to the receptor, and are thenthemselves activated by a JAK-dependent tyrosine phosphorylation event.Upon activation, STATs dissociate from the receptors, dimerize, andtranslocate to the nucleus to bind to specific DNA sites and altertranscription (Scott, M. J., C. J. Godshall, et al. (2002). “Jaks,STATs, Cytokines, and Sepsis.” Clin Diagn Lab Immunol 9(6): 1153-9).

The JAK family plays a role in the cytokine-dependent regulation ofproliferation and function of cells involved in immune response. TheJAK/STAT pathway, and in particular all four members of the JAK family,are believed to play a role in the pathogenesis of the asthmaticresponse, chronic obstructive pulmonary disease, bronchitis, and otherrelated inflammatory diseases of the lower respiratory tract. Moreover,multiple cytokines that signal through JAK kinases have been linked toinflammatory diseases or conditions of the upper respiratory tract suchas those affecting the nose and sinuses (e.g. rhinitis, sinusitis)whether classically allergic reactions or not. The JAK/STAT pathway hasalso been implicated to play a role in inflammatory diseases/conditionsof the eye including, but not limited to, iritis, uveitis, scleritis,conjunctivitis, as well as chronic allergic responses. Therefore,inhibition of JAK kinases may have a beneficial role in the therapeutictreatment of these diseases.

Blocking signal transduction at the level of the JAK kinases holdspromise for developing treatments for human cancers. Inhibition of theJAK kinases is also envisioned to have therapeutic benefits in patientssuffering from skin immune disorders such as psoriasis, and skinsensitization. Accordingly, inhibitors of Janus kinases or relatedkinases are widely sought and several publications report effectiveclasses of compounds. For example, certain JAK inhibitors, including(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile,are reported in U.S. Pat. App. Pub. No. 2007/0135461, the disclosure ofwhich is incorporated herein by reference.

In view of the growing demand for compounds for the treatment ofdisorders related to the inhibition of kinases such as Janus kinases,new and more efficient routes to inhibitors such as chiral substitutedpyrazolyl pyrrolo[2,3-d]pyrimidines and intermediates related thereto,are needed. The processes and compounds described herein help meet theseand other needs.

SUMMARY

The present invention provides, inter alia, processes of preparing acomposition comprising a compound of Formula I:

comprising reacting a compound of Formula II:

with hydrogen gas in the presence of a hydrogenation catalyst;wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   R₂ is selected from —C(═O)—NH₂, —C(═O)O—R₃, and cyano;    -   R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and    -   P₁ is a protecting group.

The present invention further provides processes of preparing acomposition comprising an enantiomeric excess of a (R)- or(S)-enantiomer of a compound of Formula I:

comprising reacting a compound of Formula II:

with hydrogen gas in the presence of a ruthenium or rhodium catalysthaving L1, wherein L₁ is a chiral phosphine ligand;wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   R₂ is selected from —C(═O)—NH₂, —C(═O)O—R₃, and cyano;    -   R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and    -   P₁ is a protecting group.

The present invention further provides processes for converting acompound of Formula I to a compound of Formula Ic, comprising reacting acompound of Formula I:

with a metal hydroxide to form a compound of Formula Ic:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   R₂ is —C(═O)O—R₃; and    -   P₁ is a protecting group.

The present invention also provides process for converting a compound ofFormula Ic to a compound of Formula Ib, comprising reacting a compoundof Formula Ic:

with ammonia or ammonium hydroxide in the presence of a coupling reagentto form a compound of Formula Ib:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   P₁ is a protecting group.

The present invention also provides processes for converting a compoundof Formula Ib to a compound of Formula Ia, comprising reacting thecompound of Formula Ib:

under dehydrating conditions to form a compound of Formula Ia:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   P₁ is a protecting group.

The present invention provide processes of preparing a compositioncomprising an enantiomeric excess of a (R)- or (S)-enantiomer of acompound of Formula Id:

comprising reacting a compound of Formula IV:

with a compound of Formula V:

in the presence of a chiral amine and an organic acid;wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   P₁ is a protecting group.

The present invention further provides processes of preparing acomposition comprising an enantiomeric excess of a (R)- or(S)-enantiomer of a compound of Formula VI:

comprising reacting a compound of Formula V:

with a compound of Formula VII:

in the presence of a chiral amine and an organic acid;wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   X₁ is halogen.

The present invention further provides a process of converting acompound of Formula VI to a compound of Formula III, comprising treatingthe compound of Formula VI:

with ammonia or ammonium hydroxide and iodine to form the compound ofFormula VIII:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   P₁ is a protecting group; and    -   X₁ is halogen.

The present invention also provides a process of converting a compoundof Formula VIII to a compound of Formula IX, comprising reacting thecompound of Formula VIII:

with a compound of Formula B-1:

to form a compound of Formula IX:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   X₁ is halo; and    -   R_(c) and R_(d) are each independently selected from H and C₁₋₆        alkyl; or    -   R_(c) and R_(d), together with the oxygen atoms to which they        are attached and the boron atom to which the oxygen atoms are        attached, form a 5- to 6-membered heterocyclic ring, which is        optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups.

The present invention also provides processes of converting a compoundof Formula IX to a compound of Formula Ia, comprising reacting thecompound of Formula IX:

with a compound of Formula X:

in the presence of a palladium catalyst and a base to form a compound ofFormula Ia:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   X₂ is a tosylate group, a triflate group, iodo, chloro, or        bromo;    -   P₁ is a protecting group; and    -   R_(c) and R_(d) are each independently selected from H and C₁₋₆        alkyl; or    -   R_(c) and R_(d), together with the oxygen atoms to which they        are attached and the boron atom to which the oxygen atoms are        attached, form a 5- to 6-membered heterocyclic ring, which is        optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups.

In some embodiments, the present invention provides compositionscomprising an enantiomeric excess of a (R)- or (S)-enantiomer of acompound of Formula IX:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   R_(c) and R_(d) are each independently C₁₋₆ alkyl; or    -   R_(c) and R_(d), together with the oxygen atoms to which they        are attached and the boron atom to which the oxygen atoms are        attached, form a 5- to 6-membered heterocyclic ring, which is        optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups.

The present invention further provides processes of preparing acomposition comprising an enantiomeric excess of a (R)- or(S)-enantiomer of a compound of Formula IX:

comprising passing a composition comprising a racemate of a compound ofFormula IX through a chiral chromatography unit using a mobile phase andcollecting a composition comprising an enantiomeric excess of the (R)-or (S)-enantiomer of a compound of Formula IX;wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   R_(c) and R_(d) are each independently C₁₋₆ alkyl; or    -   R_(c) and R_(d), together with the oxygen atoms to which they        are attached and the boron atom to which the oxygen atoms are        attached, form a 5- to 6-membered heterocyclic ring, which is        optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups.

The present invention provides processes of preparing a compositioncomprising a racemate of a compound of Formula Ia:

comprising:

a) treating a composition comprising an enantiomeric excess of the (R)-or (S)-enantiomer of a compound of Formula Ia with a compound of FormulaD-1:

in the presence of a first base under conditions sufficient to form acompound of Formula IV:

and

(b) reacting a compound of Formula IV with a compound of Formula D-1 inthe presence of a second base;

wherein:

-   -   * is a chiral carbon;    -   P₁ is a protecting group; and    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl.

The present invention further provides processes of preparing acomposition comprising a racemate of a compound of Formula Ia:

comprising treating a composition comprising an enantiomeric excess ofthe (R)- or (S)-enantiomer of a compound of Formula Ia with a compoundof Formula D-1:

in the presence of a base under conditions sufficient to form theracemate of the compound of Formula Ia;wherein:

-   -   * is a chiral carbon;    -   P₁ is a protecting group; and    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl.

The present invention further provides processes of preparing acomposition comprising an enantiomeric excess of the (R)- or(S)-enantiomer of a compound of Formula Ia:

comprising passing a composition comprising a racemate of a compound ofFormula Ia through a chiral chromatography unit using a mobile phase andcollecting a composition comprising an enantiomeric excess of the (R)-or (S)-enantiomer of a compound of Formula Ia;wherein:

-   -   * is a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   P₁ is a protecting group.

The present invention provides processes of preparing a compositioncomprising an enantiomeric excess of a (R)- or (S)-enantiomer of acompound of Formula Ia:

comprising:

(a) reacting a composition comprising a racemate of a compound ofFormula Ia with a chiral acid in the presence of a solvent to form asalt of a compound of Formula Ia;

(b) separating a composition comprising an enantiomer excess of a chiralsalt of the (R)- or (S)-enantiomer of the compound of Formula Ia; and

(c) treating the chiral salt with a base to form a compositioncomprising an enantiomeric excess of the (R)- or (S)-enantiomer of thecompound of Formula Ia;

wherein:

-   -   * is a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   P₁ is a protecting group.

The present invention further provides processes for converting acompound of Formula Ia to a compound of Formula III, comprising reactingthe compound of Formula Ia:

under deprotection conditions to form a compound of Formula III:

-   -   * is a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   P₁ is a protecting group.

The present invention further provides a process of preparing a compoundof Formula XII:

comprising reacting a compound of Formula X:

with a compound of Formula XIII:

in the presence of a palladium catalyst, base, and a solvent, to form acompound of Formula XII.wherein:

-   -   * is a chiral carbon;    -   X₂ is a tosylate group, a triflate group, iodo, chloro, or        bromo;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl; and    -   R_(c) and R_(d) are each independently H or C₁₋₆ alkyl; or    -   R_(c) and R_(d), together with the oxygen atoms to which they        are attached and the boron atom to which the oxygen atoms are        attached, form a 5- to 6-membered heterocyclic ring, which is        optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups; and    -   P₁ and P₂ are each independently a protecting group.

The present invention further provides processes for preparing acompound of Formula XVI:

comprising:

(a) reacting a compound of Formula XVIII

with about 1 or more equivalents of an C₁₋₆ alkyl Grignard reagent orC₁₋₆ alkyl lithium reagent followed by treating with about or moreequivalents of compound of Formula XVII:

and

(b) optionally, reprotecting the product of step (a) to give a compoundof Formula XVI;

wherein:

-   -   P₃ is a protecting group;    -   X₃ is halogen;    -   R₄ is C₁₋₆ alkyl; and    -   m is an integer selected from 1 and 2.

The present invention also provides a process for preparing a compoundof Formula XIa:

comprising treating a compound of Formula F-1:

with acid under conditions sufficient to form a compound of Formula XIa.

The present invention further provides compositions comprising anenantiomeric excess of a (R)- or (S)-enantiomer of a compound of FormulaI:

wherein:

-   -   * indicates a chiral carbon;    -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   R₂ is selected from —C(═O)—NH₂, —C(═O)O—R₃, —C(═O)OH, and        —C(═O)H;    -   R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and    -   P₁ is a protecting group.

The present invention also provides compounds of Formula II:

wherein:

-   -   R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆        fluoroalkyl;    -   R₂ is selected from —C(═O)—NH₂ and —C(═O)O—R₃;    -   R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and    -   P₁ is a protecting group.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedescription and drawings, and from the claims.

DETAILED DESCRIPTION

At various places in the present specification, substituents ofcompounds of the invention are disclosed in groups or in ranges. It isspecifically intended that the invention include each and everyindividual subcombination of the members of such groups and ranges. Forexample, the term “C₁₋₆ alkyl” is specifically intended to individuallydisclose methyl, ethyl, C₃ alkyl, C₄ alkyl, C₅ alkyl, and C₆ alkyl.

It is further appreciated that certain features of the invention, whichare, for clarity, described in the context of separate embodiments, canalso be provided in combination in a single embodiment. Conversely,various features of the invention which are, for brevity, described inthe context of a single embodiment, can also be provided separately orin any suitable subcombination.

The term “n-membered” where n is an integer typically describes thenumber of ring-forming atoms in a moiety where the number ofring-forming atoms is n. For example, piperidinyl is an example of a6-membered heterocycloalkyl ring and 1,2,3,4-tetrahydro-naphthalene isan example of a 10-membered cycloalkyl group.

For compounds of the invention in which a variable appears more thanonce, each variable can be a different moiety independently selectedfrom the group defining the variable. For example, where a structure isdescribed having two R groups that are simultaneously present on thesame compound, the two R groups can represent different moietiesindependently selected from the group defined for R. In another example,when an optionally multiple substituent is designated in the form:

then it is understood that substituent R can occur p number of times onthe ring, and R can be a different moiety at each occurrence. It isunderstood that each R group may replace any hydrogen atom attached to aring atom, including one or both of the (CH₂)_(n) hydrogen atoms.Further, in the above example, should the variable Q be defined toinclude hydrogens, such as when Q is the to be CH₂, NH, etc., anyfloating substituent such as R in the above example, can replace ahydrogen of the Q variable as well as a hydrogen in any othernon-variable component of the ring.

For compounds of the invention in which a variable appears more thanonce, each variable can be a different moiety independently selectedfrom the group defining the variable. For example, where a structure isdescribed having two R groups that are simultaneously present on thesame compound, the two R groups can represent different moietiesindependently selected from the group defined for R.

As used herein, the phrase “optionally substituted” means unsubstitutedor substituted. As used herein, the term “substituted” means that ahydrogen atom is removed and replaced by a substitutent. As used herein,the phrase “substituted with oxo” means that two hydrogen atoms areremoved from a carbon atom and replaced by an oxygen bound by a doublebond to the carbon atom. It is understood that substitution at a givenatom is limited by valency.

As used herein, the term “alkyl”, employed alone or in combination withother terms, refers to a saturated hydrocarbon group that may bestraight-chain or branched. In some embodiments, the alkyl groupcontains 1 to 12, 1 to 8, or 1 to 6 carbon atoms. Examples of alkylmoieties include, but are not limited to, chemical groups such asmethyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl,sec-butyl; higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl,n-hexyl, 1,2,2-trimethylpropyl, n-heptyl, n-octyl, and the like. In someembodiments, the alkyl moiety is methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl,or 2,4,4-trimethylpentyl. In some embodiments, the alkyl moiety ismethyl.

As used herein, the term “alkylcarboxamide” or “alkylaminocarbonyl”refers to a group of formula —C(O)—NH(alkyl). In some embodiments, eachalkyl group has 1 to 6 carbons.

As used herein, “alkenyl”, employed alone or in combination with otherterms, refers to an alkyl group having one or more double carbon-carbonbonds. In some embodiments, the alkenyl moiety contains 2 to 10 or 2 to6 carbon atoms. Example alkenyl groups include, but are not limited to,ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, and the like.

As used herein, “alkynyl”, employed alone or in combination with otherterms, refers to an alkyl group having one or more triple carbon-carbonbonds. Example alkynyl groups include, but are not limited to, ethynyl,propyn-1-yl, propyn-2-yl, and the like. In some embodiments, the alkynylmoiety contains 2 to 10 or 2 to 6 carbon atoms.

As used herein, the term “alkoxy”, employed alone or in combination withother terms, refers to an group of formula —O-alkyl. Example alkoxygroups include methoxy, ethoxy, propoxy (e.g., n-propoxy andisopropoxy), t-butoxy, and the like. In some embodiments, each alkylgroup has 1 to 6 carbons.

As used herein, the term “alkoxycarbonyl” refers to a group of formula—C(O)O-alkyl. In some embodiments, each alkyl group has 1 to 6 carbons.

As used herein, the term “tri-C_(n-m) alkylsilyl” refers to a group offormula —Si(alkyl)₃, wherein each alkyl group has n to m carbon atoms.In some embodiments, each alkyl group has 1 to 6 carbons.

As used herein, the term “tri-C_(n-m) alkylsilyloxy” refers to a groupof formula —OSi(alkyl)₃, wherein each alkyl group has n to m carbonatoms. In some embodiments, each alkyl group has 1 to 6 carbons.

As used herein, the term “aryl”, employed alone or in combination withother terms, refers to a monocyclic or polycyclic (e.g., having 2, 3 or4 fused rings) aromatic hydrocarbon moiety, such as, but not limited to,phenyl, 1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl, and thelike. In some embodiments, the aryl moiety may be further fused to acycloalkyl ring. In some embodiments, aryl groups have from 6 to 20carbon atoms, about 6 to 10 carbon atoms, or about 6 to 8 carbons atoms.

As used herein, the term “arylamino” refers to a group of formula—NH(aryl).

As used herein, the term “carboxy” refers to a group of formula —C(O)OH.

As used herein, the term “cycloalkyl”, employed alone or in combinationwith other terms, refers to a non-aromatic cyclic hydrocarbon moiety,which may optionally contain one or more alkenylene or alkynylene groupsas part of the ring structure. Cycloalkyl groups can include mono- orpolycyclic (e.g., having 2, 3 or 4 fused or covalently linked rings)ring systems. One or more ring-forming carbon atoms of a cycloalkylgroup can be oxidized to form carbonyl linkages. Example cycloalkylgroups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl,cycloheptatrienyl, norbornyl, norpinyl, norcarnyl, adamantyl, and thelike. In some embodiments, the cycloalkyl group is cyclopentyl.

As used herein, the term “haloalkyl”, employed alone or in combinationwith other terms, refers to an alkyl group having from one halogen atomto 2n+1 halogen atoms which may be the same or different, where “n” isthe number of carbon atoms in the alkyl group.

As used herein, the term “fluorinated alkyl”, employed alone or incombination with other terms, refers to an alkyl group having from onefluoro atom to 2n+1 fluoro atoms which may be the same or different,where “n” is the number of carbon atoms in the alkyl group. In someembodiments, the fluorinated alkyl group is trifluoromethyl.

As used herein, the terms “halo” and “halogen”, employed alone or incombination with other terms, refer to fluoro, chloro, bromo, and iodo.

As used herein, the term “heteroaryl”, “heteroaryl ring”, or “heteroarylgroup”, employed alone or in combination with other terms, refers to amonocyclic or polycyclic (e.g., having 2, 3 or 4 fused rings) aromatichydrocarbon moiety, having one or more heteroatom ring members selectedfrom nitrogen, sulfur and oxygen. In some embodiments, the heteroarylring or group has 1, 2, 3, or 4 heteratoms selected from N, O, or S. Insome embodiments, the heteroaryl ring or group has 1 or 2 rings. Whenthe heteroaryl group contains more than one heteroatom ring member, theheteroatoms may be the same or different. In some embodiments, theheteroaryl moiety may be further fused to a cycloalkyl orheterocycloalkyl ring. Examples of heteroaryl groups include withoutlimitation, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl,furyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl,pyrryl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl,pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl,isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolyl,indolinyl, and the like. In some embodiments, the heteroaryl group hasfrom 1 to about 20 carbon atoms, and in further embodiments from about 3to about 20 carbon atoms. In some embodiments, the heteroaryl groupcontains 3 to about 14, 4 to about 14, 3 to about 7, or 5 to 6ring-forming atoms. In some embodiments, the heteroaryl group has 1 toabout 4, 1 to about 3, or 1 to 2 heteroatoms. A linking heteroaryl groupis referred to herein as “heteroarylene.”

As used herein, the term “heteroarylamino” refers to a group of formula—NH(heteroaryl).

As used herein, “heterocycloalkyl” refers to non-aromatic heterocyclesincluding cyclized alkyl, alkenyl, and alkynyl groups where one or moreof the ring-forming carbon atoms is replaced by a heteroatom such as anO, N, or S atom. Heterocycloalkyl groups include monocyclic andpolycyclic (e.g., having 2, 3 or 4 fused rings) systems as well asspirocycles. Example “heterocycloalkyl” groups include morpholino,thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl,2,3-dihydrobenzofuryl, 1,3-benzodioxole, benzo-1,4-dioxane, piperidinyl,pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl,oxazolidinyl, thiazolidinyl, imidazolidinyl, and the like. Ring-formingcarbon atoms and heteroatoms of a heterocycloalkyl group can beoptionally substituted by oxo or sulfido. Also included in thedefinition of heterocycloalkyl are moieties that have one or morearomatic rings fused (i.e., having a bond in common with) to thenonaromatic heterocyclic ring, for example phthalimidyl, naphthalimidyl,and benzo derivatives of heterocycles. The heterocycloalkyl group can beattached through a ring-forming carbon atom or a ring-formingheteroatom. The heterocycloalkyl group containing a fused aromatic ringcan be attached through any ring-forming atom including a ring-formingatom of the fused aromatic ring. In some embodiments, theheterocycloalkyl group has from 1 to about 20 carbon atoms, and infurther embodiments from about 3 to about 20 carbon atoms. In someembodiments, the heterocycloalkyl group contains 3 to about 14, 4 toabout 14, 3 to about 7, or 5 to 6 ring-forming atoms. In someembodiments, the heterocycloalkyl group has 1 to about 4, 1 to about 3,or 1 to 2 heteroatoms. In some embodiments, the heterocycloalkyl groupcontains 0 to 3 double or triple bonds. In some embodiments, theheterocycloalkyl group contains 0 to 2 double or triple bonds. A linkingheterocycloalkyl group is referred to herein as “heterocycloalkylene.”

As used herein, the term “oxo” refers to a group of formula ═O.

As used herein, the term “triflate group” refers to atrifluoromethylsulfonyloxy group.

As used herein, the term “tosylate group” refers to a p-tolylsulfonyloxygroup.

The processes described herein can be monitored according to anysuitable method known in the art. For example, product formation can bemonitored by spectroscopic means, such as nuclear magnetic resonancespectroscopy (e.g., ¹H or ¹³C), infrared spectroscopy, orspectrophotometry (e.g., UV-visible); or by chromatography such as highperformance liquid chromatography (HPLC) or thin layer chromatography(TLC) or other related techniques.

As used herein, the term “reacting” is used as known in the art andgenerally refers to the bringing together of chemical reagents in such amanner so as to allow their interaction at the molecular level toachieve a chemical or physical transformation. In some embodiments, thereacting involves two reagents, wherein one or more equivalents ofsecond reagent are used with respect to the first reagent. The reactingsteps of the processes described herein can be conducted for a time andunder conditions suitable for preparing the identified product.

As used herein, the term “chiral chromatography” or “chiralchromatography column” or “chiral column” relates to a chromatographicdevice or method for separating mixtures of enantiomers or diastereomerswhich are dissolved in mobile phase. When the term “preparative” is usedin conjunction with any of the aforementioned terms, this means thedevice or method is of sufficient scale to isolate relevant quantitiesof the desired enantiomer or diastereomer. Example separation methodssuitable for chiral chromatography include HPLC (High Performance LiquidChromatography), SFC (Supercritical Fluid Chromatography), both in batchmode and in continuous mode, e.g SMB (Simulated Moving Bed), and relatedtechniques. The process of the present invention can utilize anychromatographic method for separating racemic compounds to produce theoptically pure desired enantiomer. Such methods include, but are notlimited to, traditional single column batch chromatography, continuouschromatography, or a steady state, sequential injection process (asdescribed in, for example, U.S. Pat. No. 5,630,943 and PCT Publ. No. WO98/51391). Continuous chromatographic methods include, but are notlimited to multicolumn continuous chromatographic processes, includingsuch countercurrent chromatographic processes as SMB (as described in,for example U.S. Pat. Nos. 2,985,589, 4,402,832 and 4,498,991), or anon-steady state continuous chromatographic method known as the“Varicol™” Process (as described in, for example, U.S. Pat. Nos.6,136,198; 6,375,839; 6,413,419; and 6,712,973).

In the separation of enantiomers these methods involve the use of achiral stationary phase. An achiral stationary phase may be used for theseparation of diastereomers. The term “stationary phase” relates to asuitable inert carrier material on which an interacting agent is coatedor immobilized.

As used herein, the term “chiral stationary phase” relates to stationaryphases in which the interacting agent is an enantiomerically enrichedresolving agent, for instance immobilized by coating, by chemicallybinding or by insolubilizing via cross-linking on an inert carriermaterial. A suitable inert carrier material is preferably macroporous,e.g crosslinked polystyrene, polyacrylamide, polyacrylate, alumina,kieselgur (diatomaceous), quartz, kaolin, magnesium oxide or titaniumdioxide. In some embodiments, the inert carrier material comprisessilica gel. The average particle diameter of the packing material variesdepending on the volume flow rate of the solvent flowing in thechromatographic system In some embodiments, it is 1 to 300 μm, 2 to 100μm, 5 to 75 μm or 10 to 30 μm. Appropriate selection of the averageparticle diameter of the packing material will help to adjust thepressure drop in the chromatographic process and the efficiency of thepacking material. Examples of stationary phases containing anenantiomerically enriched resolving agent are, for instance, phasesbased on either synthetic or naturally occurring chiral polymers,macrocyclic phases, ligand-exchange phases and pirkle-type phases. Suchchiral stationary phases are known and commercially available. In someembodiments, the chiral stationary phase is derivatized with at leastone sugar derivative, and in particular is a derivatized polysaccharidethat is selected from the amylosic, cellulosic, chitosan, xylan,curdlan, dextran, and inulan class of polysaccharides. In certainembodiments, the chiral stationary phase is a member of the amylosic orcellulosic class of polysaccharides. Esters and carbamates of thesematerials in particular are suitable. In additional embodiments, thechiral stationary phase is selected from cellulose phenyl carbamatederivatives, such as cellulose tris(3,5-dimethylphenyl)carbamate(available from Daicel Chemical Industries, Ltd. (Daicel) as “Chiralcel®OD” or “Chiralpak® IB”, wherein the carbamate derivative is bonded tothe cellulosic backbone); cellulose tribenzoate derivatives, such ascellulose tri 4-methylbenzoate (available from Daicel as “Chiralcel®OJ”); cellulose tricinnamate (available from Daicel as “Chiralcel® OK”);amylase phenyl and benzyl carbamate derivatives, such as amylosetris[(S)-α-methyl benzylcarbamate] (available from Daicel as “Chiralpak®AS”); amylose tris(3,5-dimethylphenyl)carbamate (available from Daicelas “Chiralpak® AD” or “Chiralpak® IA”, wherein the carbamate derivativeis bonded to the amylosic backbone); amylose 3,4-substituted phenylcarbamate or amylose 4-substituted phenyl-carbamate; and amylosetricinnamate. In some embodiments, the chiral phase is a member of thePirkle-phases family; (S,S) Whelk-O®1 and (R,R) Whelk-O®1 are preferred(available from Regis technologies Inc.).

As used herein, the term “mobile phase” relates to a solvent or mixtureof solvents in which the mixture of enantiomers to be separated isdissolved. Suitable solvents to be used in the preparativechromatographic process according to the invention are the solvents thatare known to be used in analytical chromatography. In liquidchromatography usually, non-polar, polar protic or aprotic solvents ormixture thereof are used. In supercritical chromatography preferablymixtures of carbon dioxide and polar protic solvents are used. Suitablenon polar solvents are for example hydrocarbons, for instance,n-pentane, n-hexane, hexanes, n-heptane, heptanes, cyclohexane, andmethylcyclohexane. Suitable protic or aprotic solvents are for examplealcohols, in particular methanol, ethanol, 1-propanol, 2-propanol,1-butanol, 2-butanol, isobutanol, tert butanol, ethers, for instancemethyl tert butyl ether, esters, for instance ethylacetate, halogenatedhydrocarbons and acetonitrile. The addition of water, acid (for instanceformic acid, acetic acid, trifluoroacetic acid) or base (for instanceorganic bases, e.g. trietylamine) for example less than 1% (v/v) in thesolvent may have advantageous effects.

In liquid chromatography, C₁-C₃ alcohols or mixtures of these alcoholswith hydrocarbons, for instance n-hexane or n-heptane can be used. Insupercritical chromatography mixtures of carbon dioxide and polar proticsolvents e.g. methanol, can be used. The optimal solvent (combination)can be screened using methods known in the art. A different optimalsolvent (combination) may be found when another stationary phase isused.

The compounds of the present invention also include pharmaceuticallyacceptable salts of the compounds disclosed herein. As used herein, theterm “pharmaceutically acceptable salt” refers to a salt formed by theaddition of a pharmaceutically acceptable acid or base to a compounddisclosed herein. As used herein, the phrase “pharmaceuticallyacceptable” refers to a substance that is acceptable for use inpharmaceutical applications from a toxicological perspective and doesnot adversely interact with the active ingredient. Pharmaceuticallyacceptable salts, including mono- and bi-salts, include, but are notlimited to, those derived from organic and inorganic acids such as, butnot limited to, acetic, lactic, citric, cinnamic, tartaric, succinic,fumaric, maleic, malonic, mandelic, malic, oxalic, propionic,hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, glycolic,pyruvic, methanesulfonic, ethanesulfonic, toluenesulfonic, salicylic,benzoic, and similarly known acceptable acids. Lists of suitable saltsare found in Remington's Pharmaceutical Sciences, 17th ed., MackPublishing Company, Easton, Pa., 1985, p. 1418 and Journal ofPharmaceutical Science, 66, 2 (1977), each of which is incorporatedherein by reference in their entireties.

Preparation of compounds can involve the protection and deprotection ofvarious chemical groups. The need for protection and deprotection, andthe selection of appropriate protecting groups can be readily determinedby one skilled in the art. The chemistry of protecting groups can befound, for example, in Greene, et al., Protective Groups in OrganicSynthesis, 4d. Ed., Wiley & Sons, 2007, which is incorporated herein byreference in its entirety. Adjustments to the protecting groups andformation and cleavage methods described herein may be adjusted asnecessary in light of the various substituents.

The reactions of the processes described herein can be carried out insuitable solvents which can be readily selected by one of skill in theart of organic synthesis. Suitable solvents can be substantiallynonreactive with the starting materials (reactants), the intermediates,or products at the temperatures at which the reactions are carried out,e.g., temperatures which can range from the solvent's freezingtemperature to the solvent's boiling temperature. A given reaction canbe carried out in one solvent or a mixture of more than one solvent.Depending on the particular reaction step, suitable solvents for aparticular reaction step can be selected. In some embodiments, reactionscan be carried out in the absence of solvent, such as when at least oneof the reagents is a liquid or gas.

Suitable solvents can include halogenated solvents such as carbontetrachloride, bromodichloromethane, dibromochloromethane, bromoform,chloroform, bromochloromethane, dibromomethane, butyl chloride,dichloromethane, tetrachloroethylene, trichloroethylene,1,1,1-trichloroethane, 1,1,2-trichloroethane, 1,1-dichloroethane,2-chloropropane, α,α,α-trifluorotoluene, 1,2-dichloroethane,1,2-dibromoethane, hexafluorobenzene, 1,2,4-trichlorobenzene,1,2-dichlorobenzene, chlorobenzene, fluorobenzene, mixtures thereof andthe like.

Suitable ether solvents include: dimethoxymethane, tetrahydrofuran,1,3-dioxane, 1,4-dioxane, furan, diethyl ether, ethylene glycol dimethylether, ethylene glycol diethyl ether, diethylene glycol dimethyl ether,diethylene glycol diethyl ether, triethylene glycol dimethyl ether,anisole, t-butyl methyl ether, mixtures thereof and the like.

Suitable protic solvents can include, by way of example and withoutlimitation, water, methanol, ethanol, 2-nitroethanol, 2-fluoroethanol,2,2,2-trifluoroethanol, ethylene glycol, 1-propanol, 2-propanol,2-methoxyethanol, 1-butanol, 2-butanol, i-butyl alcohol, t-butylalcohol, 2-ethoxyethanol, diethylene glycol, 1-, 2-, or 3-pentanol,neo-pentyl alcohol, t-pentyl alcohol, diethylene glycol monomethylether, diethylene glycol monoethyl ether, cyclohexanol, benzyl alcohol,phenol, or glycerol.

Suitable aprotic solvents can include, by way of example and withoutlimitation, tetrahydrofuran (THF), N,N-dimethylformamide (DMF),N,N-dimethylacetamide (DMA),1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU),1,3-dimethyl-2-imidazolidinone (DMI), N-methylpyrrolidinone (NMP),formamide, N-methylacetamide, N-methylformamide, acetonitrile, dimethylsulfoxide, propionitrile, ethyl formate, methyl acetate,hexachloroacetone, acetone, ethyl methyl ketone, ethyl acetate,sulfolane, N,N-dimethylpropionamide, tetramethylurea, nitromethane,nitrobenzene, or hexamethylphosphoramide.

Suitable hydrocarbon solvents include benzene, cyclohexane, pentane,hexane, toluene, cycloheptane, methylcyclohexane, heptane, ethylbenzene,m-, o-, or p-xylene, octane, indane, nonane, or naphthalene.

Supercritical carbon dioxide and ionic liquids can also be used assolvents.

The reactions of the processes described herein can be carried out atappropriate temperatures which can be readily determined by the skilledartisan. Reaction temperatures will depend on, for example, the meltingand boiling points of the reagents and solvent, if present; thethermodynamics of the reaction (e.g., vigorously exothermic reactionsmay need to be carried out at reduced temperatures); and the kinetics ofthe reaction (e.g., a high activation energy barrier may need elevatedtemperatures). “Elevated temperature” refers to temperatures above roomtemperature (about 22° C.).

The reactions of the processes described herein can be carried out inair or under an inert atmosphere. Typically, reactions containingreagents or products that are substantially reactive with air can becarried out using air-sensitive synthetic techniques that are well knownto the skilled artisan.

In some embodiments, preparation of compounds can involve the additionof acids or bases to affect, for example, catalysis of a desiredreaction or formation of salt forms such as acid addition salts.

Example acids can be inorganic or organic acids. Inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, andnitric acid. Organic acids include formic acid, acetic acid, propionicacid, butanoic acid, benzoic acid, 4-nitrobenzoic acid, methanesulfonicacid, p-toluenesulfonic acid, benzenesulfonic acid, tartaric acid,trifluoroacetic acid, propiolic acid, butyric acid, 2-butynoic acid,vinyl acetic acid, pentanoic acid, hexanoic acid, heptanoic acid,octanoic acid, nonanoic acid and decanoic acid.

Example bases include lithium hydroxide, sodium hydroxide, potassiumhydroxide, lithium carbonate, sodium carbonate, and potassium carbonate.Some example strong bases include, but are not limited to, hydroxide,alkoxides, metal amides, metal hydrides, metal dialkylamides andarylamines, wherein; alkoxides include lithium, sodium and potassiumsalts of methyl, ethyl and t-butyl oxides; metal amides include sodiumamide, potassium amide and lithium amide; metal hydrides include sodiumhydride, potassium hydride and lithium hydride; and metal dialkylamidesinclude sodium and potassium salts of methyl, ethyl, n-propyl, i-propyl,n-butyl, t-butyl, trimethylsilyl and cyclohexyl substituted amides.

The present invention also includes salt forms of the compoundsdescribed herein. Examples of salts (or salt forms) include, but are notlimited to, mineral or organic acid salts of basic residues such asamines, alkali or organic salts of acidic residues such as carboxylicacids, and the like. Generally, the salt forms can be prepared byreacting the free base or acid with stoichiometric amounts or with anexcess of the desired salt-forming inorganic or organic acid or base ina suitable solvent or various combinations of solvents. Lists ofsuitable salts are found in Remington's Pharmaceutical Sciences, 17thed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, the disclosureof which is hereby incorporated by reference in its entirety.

Upon carrying out preparation of compounds according to the processesdescribed herein, the usual isolation and purification operations suchas concentration, filtration, extraction, solid-phase extraction,recrystallization, chromatography, and the like may be used, to isolatethe desired products.

In some embodiments, the compounds of the invention, and salts thereof,are substantially isolated. By “substantially isolated” is meant thatthe compound is at least partially or substantially separated from theenvironment in which it was formed or detected. Partial separation caninclude, for example, a composition enriched in the compound of theinvention. Substantial separation can include compositions containing atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, at least about 95%, at least about 97%, or atleast about 99% by weight of the compound of the invention, or saltthereof. Methods for isolating compounds and their salts are routine inthe art.

Processes and Intermediates

The present invention provides, inter alia, processes of synthesizingnitrile compounds of Formula III, and intermediates thereof, which areuseful as JAK inhibitors. In one aspect, the process is a hydrogenationmethod. In some embodiments, the process is an asymmetric hydrogenationmethod, which produces an enantiomeric excess of the (R)- or(S)-enantiomer of the JAK inhibitor or intermediate thereof. In anotheraspect, the process is an asymmetric aza-Michael addition method, whichproduces an enantiomeric excess of the (R)- or (S)-enantiomer of the JAKinhibitor or intermediate thereof.

In a further aspect, the present invention provides a process forenriching the enantiomeric excess of compounds of Formula III by chiralseparation techniques or chiral salt resolution. In some embodiments,these processes involve chiral separation (such as chiral preparativechromatography) or chiral salt resolution of intermediate compounds,followed by subsequent reaction to form the compounds of Formula III. Insome embodiments, the present invention further provides a process forracemization of undesired enantiomers of intermediate compounds forproducing compounds of Formula III, which can then be resolved to givean enantiomeric excess of the desired enantiomer by the techniquesdescribed previously.

In a still further aspect, the present invention provides processes forpreparing intermediate compounds useful for producing compounds ofFormula III. In another aspect, the present invention providesintermediate compounds of any of the intermediates described herein. Instill another aspect, the present invention provides enantiomericallyenriched compositions of any of the intermediates described herein,provided the intermediates have at least one chiral center.

The processes described herein include processes for preparing compoundsand intermediates and compositions thereof, wherein R₁ is selected fromcyclopentyl, methyl and trifluoromethyl. In some embodiments, R₁ iscyclopentyl or cyclopropyl. In some embodiments, R₁ is cyclopentyl. Insome embodiments, R₁ is methyl. In some embodiments, R₁ istrifluoromethyl. These embodiments can apply to any of the intermediatesor compounds described herein in any of the processes, as appropriate.

In some embodiments, the process can be used to form a compound ofFormula III, which is3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,or pharmaceutically acceptable salt thereof. In some embodiments, theprocess can be used to form a compound of Formula III, which is(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,or pharmaceutically acceptable salt thereof. The processes describedherein are understood to include processes of preparing these compounds,especially(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile.

Processes for preparing some of the intermediates can be found in U.S.Patent Publ. No. US 20070135461, published Jun. 14, 2007 (applicationSer. No. 11/637,545, filed Dec. 12, 2006); and U.S. patent applicationSer. No. 12/138,082, filed Jun. 12, 2008, each of which is incorporatedherein by reference in its entirety.

I. Catalytic Hydrogenation Methods (Including Asymmetric HydrogenationMethods)

Compounds of Formula III can be formed by catalytic hydrogenation of acompound of Formula II to form a compound of Formula I, which can thenbe converted to a compound of Formula III through functional grouptransformation and/or deprotection steps. In some embodiments, theprocesses form a compound of Formula I as the racemate, while in morepreferred embodiments, the processes produce an enantiomeric excess ofthe (S)- or (R)-enantiomer of the compound of Formula I. One step of theprocess involves the hydrogenation of α,β-unsaturated compounds ofFormula II as shown below.

Accordingly, in one aspect, the present invention provides a process ofpreparing a composition comprising a compound of Formula I:

comprising reacting a compound of Formula II:

with hydrogen gas in the presence of a hydrogenation catalyst;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R₂ is selected from —C(═O)—NH₂, —C(═O)O—R₃, and cyano;

R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and

P₁ is a protecting group.

In some embodiments, R₁ is selected from cyclopentyl, methyl andtrifluoromethyl. In some embodiments, R₁ is cyclopentyl or cyclopropyl.In some embodiments, R₁ is cyclopentyl. In some embodiments, R₁ ismethyl. In some embodiments, R₁ is trifluoromethyl.

In some embodiments, R₂ is —C(═O)O—R₃. In some embodiments, R₂ is—C(═O)OCH₃. In some embodiments, R₂ is cyano.

In some embodiments, R₃ is selected from C₁₋₄ alkyl. In someembodiments, R₃ is selected from methyl.

The squiggly symbol for the bond connected to R₂ indicates that thecompound can be in the (E)- or (Z)-conformation. In some embodiments,when R₂ is cyano or —C(═O)—NH₂, the compound of Formula II is the(Z)-isomer, and when R₂ is —C(═O)O—R₃, the compound of Formula II is the(E)-isomer. In some embodiments, the compound of Formula II is the(Z)-isomer. In some embodiments, the compound of Formula II is the(E)-isomer.

In some embodiments, P₁ is —CH₂OC(═O)C(CH₃)₃. In some embodiments, P₁ isselected from —CH₂OCH₂CH₂Si(CH₃)₃. Appropriate P₁ protecting groupsinclude, but are not limited to the protecting groups for aminesdelineated in Wuts and Greene, Protective Groups in Organic Synthesis,4th ed., John Wiley & Sons: New Jersey, pages 696-887 (and, inparticular, pages 872-887) (2007), which is incorporated herein byreference in its entirety. In some embodiments, the protecting group forthe P₁ group is one which is stable to conditions for removing the P₂protecting group in other process steps described infra. In someembodiments, P₁ is a group which is resistant to room temperature acidicconditions. In some embodiments, P₁ is a group which is not removed infrom about 1 to about 5 N hydrochloric acid at room temperature, at atemperature from about 10° C. to about 40° C., at a temperature fromabout 15° C. to about 40° C., or at a temperature from about 15° C. toabout 30° C. In some embodiments, P₁ is benzyloxycarbonyl (Cbz),2,2,2-trichloroethoxycarbonyl (Troc), 2-(trimethylsilyl)ethoxycarbonyl(Teoc), 2-(4-trifluoromethylphenylsulfonyl)ethoxycarbonyl (Tsc),t-butoxycarbonyl (BOC), 1-adamantyloxycarbonyl (Adoc),2-adamantylcarbonyl (2-Adoc), 2,4-dimethylpent-3-yloxycarbonyl (Doc),cyclohexyloxycarbonyl (Hoc), 1,1-dimethyl-2,2,2-trichloroethoxycarbonyl(TcBOC), vinyl, 2-chloroethyl, 2-phenylsulfonylethyl, allyl, benzyl,2-nitrobenzyl, 4-nitrobenzyl, diphenyl-4-pyridylmethyl,N′,N′-dimethylhydrazinyl, methoxymethyl, t-butoxymethyl (Bum),benzyloxymethyl (BOM), or 2-tetrahydropyranyl (THP). In someembodiments, P₁ is tri(C₁₋₄ alkyl)silyl (e.g., tri(isopropyl)silyl). Insome embodiments, P₁ is 1,1-diethoxymethyl. In some embodiments, P₁ is2-(trimethylsilyl)ethoxymethyl (SEM). In some embodiments, P₁ isN-pivaloyloxymethyl (POM).

In some embodiments, the process produces a composition comprising aracemate of the compound of Formula II. Where a racemate is desired, anyhydrogenation catalyst known in the art can be utilized. In someembodiments, the hydrogenation catalyst is palladium-on-carbon.

In further embodiments, the process produces a composition comprising anenantiomeric excess of a (R)- or (S)-enantiomer of the compound ofFormula I. In general, when an enantiomeric excess of the compound ofFormula I is desired, an asymmetric hydrogenation catalyst is utilized.In some embodiments, the hydrogenation catalyst is a ruthenium orrhodium catalyst having L₁; wherein L₁ is a chiral ligand. Many suitablecatalysts are known in the art. In some embodiments, a chiral phosphineligands are used. The active catalyst systems (metal, ligand, andadditives) can be generated in situ during the reaction or generatedprior to the reaction.

In some embodiments, the catalyst can be first screened by carrying outthe catalytic asymmetric hydrogenation experiments using a relativelyhigh catalyst loading. Once the catalyst systems are selected, theexperimental conditions including the catalyst loading, hydrogenpressure, reaction solvent or solvent system, reaction temperature, andreaction time can be further optimized to improve the chemicalconversion and enentioselectivity. In some embodiments, the catalystloading is from about 0.005 to about 0.1 mole % based on the compound ofFormula II.

In some embodiments, it will be known which enantiomer of the compoundof Formula I will be produced by a particular chiral ligand. In someembodiments, the chiral ligand in the asymmetric hydrogenation catalystcan be screened to determine which enantiomer of the compound of FormulaI is produced by the process. The desired chiral ligand can then beselected so as to provide the desired enantiomer of the compound ofFormula I. For example, in some embodiments, the process furthercomprises, prior the reacting, the steps of:

(i) reacting the compound of Formula II with hydrogen gas in thepresence of a ruthenium or rhodium catalyst having L₂; and analyzing theresultant composition to determine whether the (R)- or (S)-enantiomer isin excess; wherein L₂ is a chiral ligand;

(ii) reacting the compound of Formula II with hydrogen gas in thepresence of a ruthenium or rhodium catalyst having L₃; and analyzing theresultant composition to determine whether the (R)- or (S)-enantiomer isin excess; wherein L₃ is the same chiral ligand as L₂ having theopposite stereochemistry; and

(iii) choosing L₂ or L₃ for use as L₁ based on the desiredstereochemistry for the enantiomeric excess of the composition.

In some embodiments, the hydrogenation catalyst is selected from[Ru(p-cymene)(L₁)Cl]Cl, Rh(COD)(L₁)(BF₄), Rh(COD)₂(L₁)(CF₃SO₃), andRu(L₁)(CF₃CO₂)₂. In some embodiments, the hydrogenation catalyst isselected from [Ru(L₄)(L₁)Cl]Cl, Rh(L₄)(L₁)(BF₄), Rh(L₄)₂(L₁)(CF₃SO₃),and Ru(L₁)(CF₃CO₂)₂. In some embodiments, L₄ is cumene or COD. In someembodiments, X′ is halogen. In some embodiments, X′ is chloro. In someembodiments, the hydrogenation catalyst is a mixture of [Rh(COD)₂]CF₃SO₃and a chiral phosphine ligand. In some embodiments, the solvent is2,2,2-trifluoroethanol (TFE). In some embodiments, the hydrogenationcatalyst loading is about 0.005 to about 0.01 mol %; and the ratio ofthe compound of Formula II to the hydrogenation catalyst is from about20000/1 to about 10000/1. In some embodiments, the reactionconcentration is from about 5 to about 6 mL TFE/g, the hydrogen pressureis from about 7 to about 60 bar, the reacting is run at a temperaturefrom about room temperature to about 75° C. In some embodiments, thereacting is run until the conversion of the compound of Formula II tothe compound of Formula is about equal to or greater than 99.5%. In someembodiments, the reacting is from about 10 to about 25 hours. In someembodiments, the enantiomeric excess is equal to or greater than about94%.

In some embodiments:

-   -   the hydrogenation catalyst is a mixture of [Rh(COD)₂]CF₃SO₃ and        a chiral phosphine ligand selected from:

-   -   the solvent is 2,2,2-trifluoroethanol (TFE);    -   the hydrogenation catalyst loading is about 0.005 to about 0.01        mol %;    -   the ratio of the compound of Formula II to the hydrogenation        catalyst is from about 20000/1 to about 10000/1;    -   the hydrogen pressure is from about 7 to about 60 bar;    -   the reacting is run at a temperature from about room temperature        to about 75° C.;    -   the reacting is run until the conversion of the compound of        Formula II to the compound of Formula is about equal to or        greater than 99.5%;    -   the reacting is from about 10 to about 25 hours; and    -   the enantiomeric excess is equal to or greater than about 94%.

In some embodiments, the chiral ligand is a chiral phosphine ligand. Insome embodiments, the chiral ligand is selected from one of thefollowing:

In further embodiments, the composition comprises an enantiomeric excessof the (S)-enantiomer of the compound of Formula I. In some embodiments,L₁ is selected from one of the following ligands:

In other embodiments, composition comprises an enantiomeric excess ofthe (R)-enantiomer of the compound of Formula I. In some embodiments, L₁is selected from one of the following ligands:

In some embodiments, the chiral catalyst is selected from thehydrogenation catalysts in Sigma Aldrich, “Asymmetric Catalysis:Privileged Ligands and Complexes”, ChemFiles, vol. 8, no. 2, pages 1-88,which is incorporated herein by reference in its entirety. In someembodiments, the enantiomeric excess is equal to or greater than about10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%,about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about99%, about 99.1%, about 99.2%, about 99.3%, about 99.4%, about 99.5%,about 99.6%, about 99.7%, about 99.8%, about 99.9%, or about 99.99%.

In further embodiments, the process further comprises reacting thecompound of Formula Ic under deprotection conditions to form a compoundof Formula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

In some embodiments, the process further comprises a compound of FormulaIII with phosphoric acid to form a phosphate salt of the compound ofFormula III.

Treatment of the compound of Formula Ic to remove the P₁ group can beaccomplished by methods known in the art for the removal of particularprotecting groups for amines, such as those in Wuts and Greene,Protective Groups in Organic Synthesis, 4th ed., John Wiley & Sons: NewJersey, pages 696-887 (and, in particular, pages 872-887) (2007), whichis incorporated herein by reference in its entirety. For example, insome embodiments, the P₁ group is removed by treating with fluoride ion(e.g., treating with tetrabutylammonium fluoride), hydrochloric acid,pyridinium p-toluenesulfonic acid (PPTS), or a Lewis acid (e.g., lithiumtetrafluoroborate)). In some embodiments, the treating comprisestreating with lithium tetrafluoroborate, followed by treating withammonium hydroxide (e.g., when P₁ is 2-(trimethylsilyl)ethoxymethyl). Insome embodiments, the treating comprises treating with base (e.g., P₁ isN-pivaloyloxymethyl). In some embodiments, the base is an alkali metalhydroxide. In some embodiments, the base is sodium hydroxide. In someembodiments, the treating comprises treating with sodium hydroxide orammonia in a solvent such as methanol or water.

In some embodiments, to deprotect the SEM-protection group, a mild, twostage protocol is employed. The SEM-protected substrate of Formula Ic istreated with lithium tetrafluoroborate (LiBF₄) in aqueous acetonitrileat elevated temperature, such as 80° C. for ten to twenty hours. Theresulting corresponding hydroxymethyl intermediate is then subsequentlytreated with aqueous ammonium hydroxide (NH₄OH) at room temperature toprovide the compound of Formula III.

In some embodiments, for the POM-deprotection, an aqueous sodiumhydroxide solution (NaOH) is used. Thus, a suspension of thePOM-protected compound of Formula Ic, is treated with a 1 N aqueoussodium hydroxide solution at room temperature for two to three hours.The desired product of Formula III can be obtained after the typicalacid-base work-up. In some embodiments, the deprotecting conditionscomprise treating with lithium tetrafluoroborate, followed by treatingwith aqueous ammonium hydroxide.

In some embodiments, the process further comprises reacting a compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

Processes of Converting the Amide of Formula I to a Nitrile of FormulaIII

The present invention further provides a process of converting an amideof Formula I to a nitrile compound of Formula I. The methods ofconverting the amide of Formula I involve dehydrating the amide to forma nitrile. The protecting group can then be removed and the resultantamine can be protonated to form a pharmaceutically acceptable salt.Accordingly, in some embodiments, the present invention provides aprocess comprising reacting a compound of Formula I:

under dehydrating conditions to form a compound of Formula Ia:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R₂ is selected from —C(═O)—NH₂;

P₁ is a protecting group.

In some embodiments, the dehydrating conditions comprise trichloroacetylchloride in the presence of triethylamine. In some embodiments, thedehydrating conditions comprise any dehydrating agent for dehydration ofamides, including but not limited to, an acid chloride (e.g.,trichloroacetyl chloride), P₂O₅; ZnCl₂ (under microwave conditions);triphenylphosphine and N-chlorosuccinimide; ethyl dichlorophosphate/DBU;and PdCl₂. In some embodiments, the dehydrating conditions are thosedescribed in Kuo, C-W.; Zhu, J.-L.; Wu, J.; et al. Chem. Commun. 2007,301; Manjula, K.; Pasha, M. A. Syn. Commun. 2007, 37, 1545; Takahashi,T.; Sugimoto, O.; Koshio, J.; Tanji, K. Heterocycles 2006, 68, 1973;Maffioli, S. I.; Marzorati, E.; Marazzi, A. Organic Letters 2005, 7,5237; or Iranpoor, N.; Firouzabadi, H.; Aghapour, G. Syn. Commun. 2002,32, 2535, each of which is incorporated by reference in its entirety.

In further embodiments, the process further comprises reacting thecompound of Formula Ic under deprotection conditions to form a compoundof Formula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

Appropriate P₁ groups and deprotection methods include, but are notlimited to those described supra.

In some embodiments, the process further comprises reacting a compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

Processes of Converting the Ester of Formula I to a Nitrile of FormulaIII

The present invention further provides a process of converting an esterof Formula I to a nitrile compound of Formula I. The processes ofconverting the ester of Formula I involves saponification of the esterto form an acid, selective ammonlysis, and dehydration of the amide. Theprotecting group can then be removed and the resultant amine can beprotonated to form a pharmaceutically acceptable salt.

Accordingly, the present invention provides a process comprisingreacting the compound of Formula I:

with a metal hydroxide to form a compound of Formula Ic:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R₂ is —C(═O)OR₃;

R₃ is selected from C₁₋₄ alkyl; and

P₁ is a protecting group.

In some embodiments, the metal hydroxide is an alkali metal hydroxide oran alkaline earth hydroxide. In some embodiments, the metal hydroxide islithium hydroxide.

In further embodiments, the process further comprises reacting thecompound of Formula Ic with ammonia or ammonium hydroxide in thepresence of a coupling reagent to form a compound of Formula Ib:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

In some embodiments, the coupling agent is N,N-carbonyldiimidazole. Insome embodiments, the coupling agent is selected from1,2-benzisoxazol-3-yl diphenyl phosphate; ClCO₂-i-Bu and Et₃N;carbodiimide; SOCl₂ and Cl—C(O)—C(O)—Cl; tosyl chloride and DMAP; andClCO₂-i-Bu and triethylamine. In some embodiments, the coupling agent isselected from those in: Ueda, M.; Oikawa, H. J. Org. Chem. 1985, 50,760. (1,2-benzisoxazol-3-yl diphenyl phosphate); Lai, M.; Liu, H. J. Am.Chem. Soc. 1991, 113, 7388. (ClCO₂-i-Bu, Et₃N); Williams, A.; Ibrahim,I. Chem. Rev. 1991, 81, 589. (Carbodiimide); Weiss, M. M.; Harmange, J;Polverino, A. J. J. Med. Chem., 2008, 51, 1668. (SOCl2, Cl—CO—CO—Cl);Hong, C. Y.; and Kishi. Y. J. Am. Chem. Soc., 1991, 113, 9693. (TsCl,DMAP); and Nitta, H.; Yu, D.; Kudo, M.; Mori, A.; Inoue, S. J. Am. Chem.Soc., 1992, 114, 7969. (ClCO₂-i-Bu, Et₃N).

In other embodiments, the process further comprises reacting thecompound of Formula Ib under dehydrating conditions to form a compoundof Formula Ia:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

In some embodiments, the dehydrating conditions comprise trichloroacetylchloride in the presence of triethylamine. In some embodiments, thedehydrating conditions comprise any dehydrating agent for dehydration ofamides, including but not limited to, an acid chloride (e.g.,trichloroacetyl chloride), P₂O₅; ZnCl₂ (under microwave conditions);triphenylphosphine and N-chlorosuccinimide; ethyl dichlorophosphate/DBU;and PdCl₂. In some embodiments, the dehydrating conditions are thosedescribed in Kuo, C-W.; Zhu, J.-L.; Wu, J.; et al. Chem. Commun. 2007,301; Manjula, K.; Pasha, M. A. Syn. Commun. 2007, 37, 1545; Takahashi,T.; Sugimoto, O.; Koshio, J.; Tanji, K. Heterocycles 2006, 68, 1973;Maffioli, S. I.; Marzorati, E.; Marazzi, A. Organic Letters 2005, 7,5237; or Iranpoor, N.; Firouzabadi, H.; Aghapour, G. Syn. Commun. 2002,32, 2535, each of which is incorporated by reference in its entirety.

In further embodiments, the process further comprises reacting thecompound of Formula Ic under deprotection conditions to form a compoundof Formula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

Appropriate P₁ groups and deprotection methods include, but are notlimited to those described supra.

In some embodiments, the process further comprises reacting a compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

Starting Materials for the Hydrogenation Processes (Compounds of FormulaII)

The compounds of Formula II, used in the asymmetric hydrogenationprocesses (supra), can be made as shown in the Scheme 1, wherein P₁ andP₂ are each, independently, a protecting group, R₁ is selected from C₃₋₇cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl, and R₂ is cyano or analkyl ester. The routes for preparing compounds of Formula IV aredescribed infra.

The process involves an aza-Michael addition reaction between anappropriately substituted acetylene of Formula XIV with a protected4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidine compound of Formula IV(preparation of compounds of Formula IV and XIV are described infra).This reaction can be conducted under the influence of catalytic amountof solid potassium carbonate in DMF at room temperature to afford thecorresponding compound of Formula I.

Compounds of Formula II, wherein R₁ is —C(═O)NH₂, can be formed as shownin Scheme 2, by treating a compound of Formula IIa with an acid to forma compound of Formula IIb.

Accordingly, the present invention provides a method of preparing acompound of Formula II:

comprising reacting a compound of Formula IV:

with a compound of Formula XIV:

in the presence of a base;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R₂ is selected from —C(═O)O—R₃ and cyano;

R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and

P₁ is a protecting group.

Appropriate P₁ protecting groups include, but are not limited to, thoselisted supra.

In some embodiments, the aza-Michael addition is conducted in an organicsolvent at room temperature in the presence of a catalytic amound ofbase. The base can be suitable solvent or base for aza-Michaelreactions. In some embodiments, the solvent is acetonitrile ordimethylformide (DMF). In some embodiments, the base is atetraalkylammonium halide, tetraalkylammonium hydroxide, guanidine,amidine, hydroxide, alkoxide, silicate, alkali metal phosphate, oxide,tertiary amine, alkali metal carbonate, alkali metal bicarbonate, alkalimetal hydrogen phosphate, phosphine, or alkali metal salt of acarboxylic acid. In some embodiments, the base is tetramethyl guanidine,1,8-diazabicyclo(5.4.0)undec-7-ene, 1,5-diazabicyclo(4.3.0)non-5-ene,1,4-diazabicyclo(2.2.2)octane, tert-butyl ammonium hydroxide, sodiumhydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide,tripotassium phosphate, sodium silicate, calcium oxide, triethylamine,sodium carbonate, potassium carbonate, sodium bicarbonate, potassiumbicarbonate, potassium hydrogen phosphate, triphenyl phosphine, triethylphosphine, potassium acetate, or potassium acrylate. In someembodiments, the base is 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) orpotassium carbonate. In some embodiments, the base is DBU. In someembodiments, the base is present in a catalytic amount. In someembodiments, the amount of base is about 0.1 to about 5 equivalents, orabout 0.5 to about 3 equivalents, or about 0.1 to about 0.5 equivalents.In some embodiments, the reaction is complete in about 1 to about 3hours.

In some embodiments, R₁ is selected from cyclopentyl, methyl andtrifluoromethyl. In some embodiments, R₁ is cyclopentyl or cyclopropyl.In some embodiments, R₁ is cyclopentyl. In some embodiments, R₁ ismethyl. In some embodiments, R₁ is trifluoromethyl.

In some embodiments, the base is an alkali metal or alkaline earth metalcarbonate. In some embodiments, the base is potassium carbonate.

In some embodiments, the present invention provides a compound ofFormula II:

wherein:

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R₂ is selected from —C(═O)—NH₂ and —C(═O)O—R₃;

R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and

P₁ is a protecting group.

In some embodiments, P₁ is —CH₂C(═O)C(CH₃)₃ or —CH₂OCH₂CH₂Si(CH₃)₃. Insome embodiments, R₁ is cyclopentyl.

Compounds of Formula IIb, wherein R₁ is —C(═O)NH₂, can be formed bytreating a compound of Formula IIa:

with an acid to form a racemic form of a compound of IIb:

wherein:

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

In some embodiments, the acid is trifluoroacetic acid, sulfuric acid, ora combination thereof. In some embodiments, the treating comprisestreating with trifluoroacetic acid (TFA) and sulfuric acid (H₂SO₄) atroom temperature. In some embodiments, the ratio of TFA to H₂SO₄ isabout 10:1 by volume. In some embodiments, the reaction is completewithin about one hour.

Compounds of Formula XIV, used in the process described in Scheme 1, canbe formed by methods such as those shown in Scheme 3 below. Accordingly,a compound of Formula XIVa (wherein R₂ of Formula XIV is cyano) isprepared by treating the lithium salt of a compound of Formula C-1 withcyanatobenzene (C-2), which is in situ generated from phenol and cyanicbromide, in an organic solvent, such as anhydrous THF, at about −78° C.to about room temperature to afford the corresponding 3-substitutedpropiolonitrile of Formula XIVa. Similarly, the lithium salt of acompound of Formula C-1 treated with an chloroformate of Formula C-3provides a 3-substituted propiolate compound of Formula XIVb (wherein R₂of Formula XIV is —C(═O)OR₃).

II. Asymmetric Aza-Michael Addition Processes for Preparing an AldehydeIntermediate of Formula Id or VI

In another aspect, the present invention provides, inter alia, anenantiomeric excess of a (R)- or (S)-enantiomer of a compound of FormulaId:

comprising reacting a compound of Formula IV:

with a compound of Formula V:

in the presence of a chiral amine and an organic acid;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

P₁ is a protecting group.

In a further aspect, the present invention provides a method ofpreparing an enantiomeric excess of a (R)- or (S)-enantiomer of acompound of Formula VI:

comprising reacting a compound of Formula V:

with a compound of Formula VII:

in the presence of a chiral amine and an organic acid;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

X₁ is halogen.

While not wishing to be bound by any particular theory, the mechanism ofthese chiral amine catalyzed aza-Michael conjugate addition ofN-heterocyclic compounds to α,β-unsaturated aldehydes is understood toinvolve the following pathways. First, the α,β-unsaturated aldehyde ofFormula V reacts with the protonated catalyst formed from thecombination of the chiral amine and the organic acid and forms animinium ion with loss of water. Owing to the chirality of the catalyst,two different iminium ions that have E and Z configurations can beformed. The corresponding E configuration is expected to be the majorintermediate present in which the Si face is shielded by the chiralgroup in the catalyst, leaving Re face available for the approach of theN-heterocyclic compounds. Second, the addition of substituted pyrazoleto the iminium ion gives the enamine intermediate, which bears apositive charge on the protonated pyrazole ring. This proton is thentransferred from the nitrogen atom in the pyrazole ring to the enaminecarbon atom to form iminium intermediate. Third, the hydrolysis of theiminium ion leads to regeneration of the catalyst and product. Based onthe understanding of reaction mechanism, the reaction conditions forthis organocatalyzed aza-Michael reaction were defined.

In some embodiments, the compound of Formula V is present in excessamounts (e.g., from about 1.5 to about 5 equivalents). In someembodiments, the chiral amine is present in about 0.02 to about 0.15equivalents, or about 0.05 to about 0.10 equivalents.

In some embodiments of either asymmetric aza-Michael addition process,the organic acid is p-toluenesulfonic acid, benzoic acid or4-nitrobenzoic acid. In some embodiments, the organic acid is benzoicacid. In some embodiments, organic acid is present in about 0.05 toabout 0.10 equivalents.

In some embodiments, the reacting is conducted in an organic solventselected from chloroform (CHCl₃) or toluene. In some embodiments, thereacting is at a temperature of about room temperature, or from about 0to about 5° C. In some embodiments, the reaction is complete in about 10to about 24 hours. In some embodiments, the reaction conversion reachesover 95% with the isolated yield to about 80 to about 90%. Chiral HPLCmethods have been developed to determine the chiral purity of eachaza-Michael adduct or its derivative.

In some embodiments of either asymmetric aza-Michael addition process,the chiral amine is a (R)- or (S)-enantiomer of a compound of FormulaA-1:

wherein:

X is CY₃Y₄ and Y is CY₅Y₆; or

X is S or NY₇ and Y is CY₅Y₆; or

X is CY₃Y₄ and Y is S;

Q₁ and Q₂ are each independently selected from H, C₁₋₆ alkyl, C₁₋₆haloalkyl, carboxy, C₁₋₆ alkylcarboxamide, C₁₋₆ alkoxycarbonyl, andphenyl; wherein the C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkylcarboxamide,C₁₋₆ alkoxycarbonyl, and phenyl are each optionally substituted by 1, 2,or 3 groups independently selected from hydroxyl, carboxy, tri-C₁₋₆alkylsilyl, tri-C₁₋₆ alkylsilyloxy, C₆₋₁₀ aryl, C₆₋₁₀ arylamino, C₁₋₉heteroaryl, and C₁₀ heteroarylamino; wherein the C₆₋₁₀ aryl, C₆₋₁₀arylamino, C₁₋₉ heteroaryl, and C₁₋₉ heteroarylamino are each optionallysubstituted by 1, 2, 3, or 4 groups independently selected from halogen,C₁₋₆ alkyl, and C₁₋₆ haloalkyl; and

Y₁, Y₂, Y₃, Y₄, Y₅, Y₆ are each independently selected from H, hydroxyl,carboxy, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxycarbonyl, and phenyl; or

Y₁ and Y₂ together form oxo; or

Y₃ and Y₄ together form oxo; or

Y₅ and Y₆ together form oxo; or

Y₁ and Y₂, together with the carbon to which they are attached, form a5- or 6-membered cycloalkyl ring; or

Q₁ and Y₅, together with the carbon atoms to which they are attached,form a 5- or 6-membered cycloalkyl ring.

In some embodiments of the compounds of Formula A-1:

X is CY₃Y₄ and Y is CY₅Y₆; or

X is S or NY₇ and Y is CY₅Y₆; or

X is CY₃Y₄ and Y is S;

Q₁ is H or methyl;

Q₂ is selected from H, methyl, isopropyl, butyl, carboxy, C₁₋₅alkylaminocarbonyl, methoxycarbonyl, and phenyl; wherein the methyl andC₁₋₅ alkylaminocarbonyl are each optionally substituted by 1, 2, or 3groups independently selected from hydroxyl, carboxy, tri-C₁₋₆alkylsilyl, tri-C₁₋₄ alkylsilyloxy, phenyl, phenylamino, and indol-3-yl;wherein the phenyl and the indol-3-yl are each optionally substituted by1 or 2 groups independently selected from methyl and trifluormethyl;

Y₁ is H, hydroxyl, carboxy, methyl, and methoxycarbonyl;

Y₂ is H or methyl;

Y₃, Y₄, Y₅, and Y₆ are each independently selected from H, hydroxyl,methyl, and phenyl;

Y₇ is H or methyl; or

Y₁ and Y₂ together form oxo; or

Y₃ and Y₄ together form oxo; or

Y₅ and Y₆ together form oxo; or

Y₁ and Y₂, together with the carbon to which they are attached, form a6-membered cycloalkyl ring; or

Q₁ and Y₅, together with the carbon atoms to which they are attached,form a 6-membered cycloalkyl ring.

In some embodiments of either asymmetric aza-Michael addition process,the chiral amine is a (R)- or (S)-enantiomer of a compound of FormulaA-2:

wherein

* is a chiral carbon having a (R)- or (S)-configuration;

Ar₁ and Ar₂ are each independently C₆₋₁₀ aryl, which is optionallysubstituted by 1, 2, 3, or 4 groups independently selected from C₁₋₆alkyl and C₁₋₆ haloalkyl;

each R_(a) are independently selected from C₁₋₆ alkyl; and

R_(b) is selected from H, C₁₋₆ alkyl, and C₁₋₆ haloalkyl.

In some embodiments, Ar₁ and Ar₂ are each independently phenyl, which isoptionally substituted by 1, 2, 3, or 4 groups independently selectedfrom methyl and trifluoromethyl; each R_(a) is independently selectedfrom methyl, ethyl, or t-butyl; and R_(b) is H.

In some embodiments of either asymmetric aza-Michael addition process,the chiral amine is a (R)- or (S)-enantiomer of a compound selected fromproline, prolinamide, prolyl-L-leucine, prolyl-L-alanine, prolylglycine,prolyl-L-phenylalanine, diphenylpyrrolidine, dibenzylpyrrolidine,N-(1-methylethyl)-pyrrolidinecarboxamide, 2-(anilinomethyl)pyrrolidine,2-[bis(3,5-dimethylphenyl)methyl]pyrrolidine,diphenyl(pyrrolidin-2-yl)methanol, prolinol, 4-thiazolidinecarboxylicacid, trans-3-hydroxyproline, trans-4-hydroxyproline,4-benzyl-1-methyl-imidazolidine-2-carboxylic acid,1-methyl-4-phenyl-imidazolidine-2-carboxylic acid,4,5-octahydro-benzoimidazole-2-carboxylic acid,4,5-diphenyl-imidazolidine-2-carboxylic acid,N1-methyl-3-phenylpropane-1,2-diamine, 1,2-diphenylethanediamine,1-methyl-4-(1-methyl-1H-indol-3-ylmethyl)-imidazolidine-2-carboxylicacid, 4-benzyl-1-methyl-imidazolidine-2-carboxylic acid,1,2-cyclohexanediamine, 2-phenyl-thiazolidine-4-carboxylic acid,tert-leucine methyl ester, 5-benzyl-2,2,3-trimethyl-imidazoline-4-one,methyl prolinate, 4,5-diphenylimidazolidine,2-cyclohexyl-4,5-diphenylimidazolidine,2-{bis-[3,5-bis(trifluoromethyl)phenyl]-trimethylsilanyloxy-methyl}-pyrrolidine,2-{bis-[3,5-dimethylphenyl]-trimethylsilanyloxy-methyl}-pyrrolidine,2-{diphenyl-trimethylsilanyloxy-methyl}-pyrrolidine,2-{bis[naphth-2-yl]-trimethylsilanyloxy-methyl}-pyrrolidine,2-{tert-butyldimethylsilyloxy-diphenyl-methyl}-pyrrolidine,2-{bis-[3,5-bis(trifluoromethyl)phenyl]-triethylsilanyloxy-methyl}-pyrrolidine,and2-{bis-[3,5-bis(trifluoromethyl)phenyl]-ethyl-dimethylsilyloxy-methyl}-pyrrolidine;wherein the (R)- or (S)-configuration is at the carbon adjacent to a NHgroup in the compound.

In some of the preceding embodiments, the chiral amine is the(R)-enantiomer.

In some embodiments of either asymmetric aza-Michael addition process,the chiral amine is selected from one of the following compounds:

In some embodiments, the enantiomeric excess is from about 85% to about95%. In some embodiments, the enantiomeric excess is equal to or greaterthan about 10%, about 20%, about 30%, about 40%, about 50%, about 60%,about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about98%, about 99%, about 99.1%, about 99.2%, about 99.3%, about 99.4%,about 99.5%, about 99.6%, about 99.7%, about 99.8%, about 99.9%, orabout 99.99%.

In some embodiments, the present invention provides a compositioncomprising an enantiomeric excess of a (R)- or (S)-enantiomer of acompound of Formula I:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R₂ is selected from —C(═O)—NH₂, —C(═O)O—R₃, —C(═O)OH, and —C(═O)H;

R₃ is selected from C₁₋₄ alkyl or C₁₋₄ fluoroalkyl; and

P₁ is a protecting group.

In some embodiments, P₁ is —CH₂C(═O)C(CH₃)₃ or —CH₂OCH₂CH₂Si(CH₃)₃. Insome embodiments, R₁ is cyclopentyl.

In other embodiments, the present invention provides a compositioncomprising an enantiomeric excess of a (R)- or (S)-enantiomer of acompound of Formula IX:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

R_(c) and R_(d) are each independently C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups.

In some embodiments, R₁ is cyclopentyl.

Processes for Converting the Aldehyde Intermediates of Formula I or VIto a Nitrile Compound

In another aspect, the present invention provides a process forpreparing a nitrile compound from a compound of Formula Id. Accordingly,in some embodiments, the present invention provides a process comprisingtreating the compound of Formula Id:

with ammonia or ammonium hydroxide and iodine to form the compound ofFormula Ia:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

In some embodiments, the treating is accomplished by treatment of thechiral aldehyde of Formula I with excess amount of aqueous ammonium(NH₄OH) and stoichiometric amount of iodine (I₂) in an organic solvent,such tetrahydrofuran (THF), at room temperature. In some embodiments,the reaction is complete within about 1 to about 2 hours at roomtemperature. The chirality of the chiral aldehydes is kept intact undersuch reaction conditions. The chirality of the chiral nitriles can bechecked by chiral HPLC analysis.

In some embodiments, the process further comprises reacting the compoundof Formula Ic under deprotection conditions to form a compound ofFormula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

Appropriate P₁ groups include, but are not limited to those describedsupra.

In some embodiments, the process further comprises reacting the compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

In a further aspect, the present invention provides a process forpreparing a nitrile compound from a compound of Formula VI. Accordingly,in some embodiments, the present invention provides a process comprisingtreating the compound of Formula VI:

with ammonia or ammonium hydroxide and iodine to form the compound ofFormula VIII:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

X₁ is halogen.

In some embodiments, the process further comprises reacting the compoundof Formula VIII with a compound of Formula B-1:

to form a compound of Formula IX:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

R_(c) and R_(d) are each independently selected from H and C₁₋₆ alkyl;or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups.

In some embodiments, the compound of Formula B-1 is4,4,5,5,4′,4′,5′,5′-octamethyl-[2,2′]bis[1,3,2-dioxaborolanyl].

In further embodiments, the process further comprises reacting thecompound of Formula IX with a compound of Formula X:

in the presence of a palladium catalyst and a base to form a compound ofFormula Ic:

wherein

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R_(c) and R_(d) are each independently selected from H and C₁₋₆ alkyl;or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups;

X₂ is a tosylate group, a triflate group, iodo, chloro, or bromo; and

P₁ is a protecting group.

In further embodiments, the process further comprises reacting acompound of Formula IX with a compound of Formula XI:

in the presence of a palladium catalyst, base, and a solvent, to form acompound of Formula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R_(c) and R_(d) are each independently selected from H and C₁₋₆ alkyl;or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups; and

X₂ is a tosylate group, a triflate group, iodo, chloro, or bromo.

In some embodiments, X₂ is bromo, iodo, or chloro. In some embodiments,X₂ is chloro.

The Suzuki coupling reaction can be initiated using a number ofpalladium(0) and palladium(II) catalysts and performed under conditionsknown in the art (see, e.g., Miyaura and Suzuki, Chem. Rev. 1995, 95,2457-2483, which is hereby incorporated in its entirety). In someembodiments, the palladium catalyst is Pd(PPh₃)₄ and Pd(dppf)₂Cl₂.

In some embodiments, the palladium catalyst istetrakis(triphenylphosphine)palladium(0) ortetrakis(tri(o-tolyl)phosphine)palladium(0). In some embodiments, thepalladium catalyst is tetrakis(triphenylphosphine)palladium(0).

In some embodiments, the palladium catalyst loading is from about 1×10⁻⁴to about 0.1 equivalents. In some embodiments, the palladium catalystloading is from about 0.0010 to about 0.0015 equivalents. In someembodiments, the stoichiometric ratio of the compound of Formula X or XIto the compound of Formula IX is from about 1:1.05 to about 1:1.35.

In some embodiments, the solvent comprises water and an organic solvent.In some embodiments, the organic solvent is 1,4-dioxane, 1-butanol,1,2-dimethoxyethane (DME), 2-propanol, toluene or ethanol, or acombination thereof. In some embodiments, the organic solvent comprisesDME. In some embodiments, the organic solvent comprises DMF.

In some embodiments, the base is an inorganic base. In some embodiments,the base is an organic base. In some embodiments, the base is an alkalimetal carbonate. In some embodiments, the base is potassium carbonate(K₂CO₃). In some embodiments, two to five equivalents of base (e.g.,K₂CO₃) are used.

In some embodiments, the Suzuki coupling reaction is conducted at atemperature of about 80 to about 100° C. In some embodiments, thereaction is carried out for two to twelve hours. In some embodiments,the compound of Formula XII can be optionally isolated from aqueouswork-up of the Suzuki coupling reaction mixture or directly used.Appropriate P₂ protecting groups include, but are not limited to theprotecting groups for amines delineated in Wuts and Greene, ProtectiveGroups in Organic Synthesis, 4th ed., John Wiley & Sons: New Jersey,pages 696-887 (and, in particular, pages 872-887) (2007), which isincorporated herein by reference in its entirety.

In other embodiments, the process further comprises reacting thecompound of Formula Ia under deprotection conditions to form a compoundof Formula III:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

Appropriate P₁ groups and deprotection methods include, but are notlimited to those described supra.

In some embodiments, the process further comprises reacting a compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

Starting Materials for the Aza-Michael Addition Processes

A compound of Formula IV can be formed by methods analogous to thosedescribed infra. 3-Substituted acrylaldehydes of Formula V can, in turn,be prepared as shown in Scheme 4. Accordingly, treatment of an aldehydeof Formula C-4 under typical Wittig conditions (e.g., reaction with(triphenylphosphoranylidene)acetaldehyde) provides the correspondingcompound of Formula V.

III. Synthesis and Racemic Resolution of Pyrazole Intermediates

Chiral compounds of Formula III can be produced by chiral columnseparation (such as by chiral preparative chromatography) of a racemateof a protected pyrazole borate derivative of Formula IX, followed by aSuzuki coupling reaction of the chiral intermediate of IX with aunprotected pyrrolo[2,3-d]pyrimidine of Formula XI (Scheme 5).Alternatively, the chiral intermediate of Formula (S)-IX or (R)-IX canbe reacted under Suzuki coupling conditions with a protectedpyrrolo[2,3-d]pyrimidine of Formula X, followed by deprotection toremove the P₁ protecting group to give a chiral compound of Formula III(Scheme 5). The racemic substituted pyrazole borate derivatives ofFormula IX can be produced via the Michael addition reaction betweenpyrazole boronic derivative of Formula XV and a Michael acceptor ofFormula D-1 (Scheme 5).

Accordingly, in some embodiments, the present invention provides aprocess of preparing a composition comprising an enantiomeric excess ofthe (R)- or (S)-enantiomer of a compound of Formula IX:

comprising passing a composition comprising a racemate of a compound ofFormula IX through a chiral chromatography unit using a mobile phase andcollecting a composition comprising an enantiomeric excess of the (R)-or (S)-enantiomer of a compound of Formula IX;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R_(c) and R_(d) are each independently C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups. In some embodiments, thechromatography is carried out in either batch or continuous mode using achiral stationary phase and a mobile phase in isocratic or gradientmode.

In some embodiments, the chiral chromatography unit is a preparativehigh performance liquid chromatography (HPLC) system equipped with achiral column, which is packed with a chiral stationary phase. In someembodiments, the chiral column is packed with a chiral stationary phasecomprising amylose tris(3,5-dimethylphenyl carbamate immobilized onsilica gel (available from Daicel as “Chiralpak® IA”). In someembodiments, the chiral column is packed with a chiral stationary phasecomprising cellulose tris(3,5-dimethylphenyl carbamate) coated on silicagel (available from Daicel as “Chiralcel® ChiralcelOD”). In someembodiments, the chromatography unit is a continuous chromatographyprocess such as simulated moving bed (SMB) chromatography or Varicolprocess using a unit equipped with a set of eight columns each packedwith a chiral stationary phase. In some embodiments the unit is equippedwith 3 to 12 columns, or 5 to 10 columns, or 5 to 8 columns, each packedwith a chiral stationary phase, in some instances, the same chiralstationary phase. In some embodiments, the column is packed with chiralstationary phase made of amylose tris(3,5-dimethylphenyl carbamate)immobilized on silica gel (available from Daicel as “Chiralpak® IA). Insome embodiments, the column is packed with a chiral stationary phasemade of cellulose tris(3,5-dimethylphenyl carbamate) coated on silicagel (available from Daicel as “Chiralcel® OD”). In some embodiments, thechiral stationary phase is cellulose modified chiral stationary phase(CSP, Chiral Technologies. In some embodiments, the chiral stationaryphase is a silica gel based stationary phase coated with 4-(3,5-dinitrobenzamido)tetrahydrophenanthrene (available from Regis Technologies as“(S,S) Whelk-O® 1”). In some embodiments, the mobile phase comprisesethanol and hexanes. In some embodiments, the mobile phase comprisesabout 1:9 ratio of ethanol to hexanes. In some embodiment the hexanesare replaced by heptanes, n-heptane, cyclohexane or methylcyclohexane.In some embodiments, the ethanol is present in an amount of about 10% toabout 100% by volume, or about 10% to about 25% by volume, or about 15%ethanol. In some embodiments, the mobile phase comprises about 15%ethanol and about 85% hexanes by volume. In some embodiments, the mobilephase comprise ethanol and hexanes, wherein the ethanol is present in anamount of about 25% to about 10% by volume. In some embodiments, themobile phase comprises isopropanol and hexanes, wherein the isopropanolis present in an amount of about 25% to about 10% by volume. In someembodiments, the hexanes are replaced by heptanes, n-heptane,cyclohexane or methylcyclohexane. In some embodiments, the isopropanolis present in an amount of about 10% to about 25% by volume. In someembodiments, the mobile phase comprises methyl-tert-butyl ether andhexanes. In some embodiments the hexanes are replaced by heptanes,n-heptane, cyclohexane or methylcyclohexane. In some embodiment themethyl-tert-butyl ether is present in an amount of about 10% to about100% by volume, preferably about 50% to about 100% by volume, and mostpreferably about 90% to about 100% by volume. In some embodiments, themobile phase comprises ethyl acetate and hexanes. In some embodiments,the hexanes are replaced by heptanes, n-heptane, cyclohexane ormethylcyclohexane. In some embodiments, the ethyl acetate is present inan amount of about 10% to about 100% by volume, about 50% to about 100%by volume, or about 75% by volume. In some embodiments, the mobile phasecomprises tetrahydrofuran and hexanes. In some embodiments, the hexanesare replaced by heptanes, n-heptane, cyclohexane or methylcyclohexane.In some embodiments, the tetrahydrofuran is present in an amount ofabout 10% to about 100% by volume, about 10% to about 50% by volume, orabout 25% by volume. In some embodiments, the chromatography unit iskept at room temperature. In some embodiments, the mobile phase ispassed at a flow rate of about 1 mL per minute to about 20 mL perminute. In some embodiments, the mobile phase is passed at a flow rateof about 1 mL per minute. In some embodiments, the mobile phase ispassed at a flow rate of about 18 mL per minute. In some embodiments,the eluent is monitored by ultraviolet (UV) spectroscopy. In someembodiments, the eluent is monitored by ultraviolet spectroscopy atabout 220 nm. Collection of the portion of the eluent containing theenantiomerically enriched composition can be determined by detection ofthe elution of the desired enantiomer by UV spectroscopy. Determinationof the % ee (enantiomeric excess) of the composition can then bedetermined by analytical chiral HPLC.

In some embodiments, the chromatographic method employed is batchpreparative chromatography, supercritical fluide chromatography (SFC), acyclojet process, a continuous multicolumn chromatography process, asimulated moving bed process, a Varicol™ process, or a PowerFeedprocess.

In some embodiments, the chiral stationary phase comprises aninteracting agent which is an enantiomerically enriched resolving agent,immobilized to an inert carrier material by, for example, chemicallybinding or by insolubilizing via cross-linking. The suitable inertcarrier material can be macroporous, e.g crosslinked polystyrene,polyacrylamide, polyacrylate, alumina, kieselgur (diatomaceous), quartz,kaolin, magnesium oxide, titanium dioxide or silica gel. In someembodiments, the inert carrier material is Silicagel.

In some embodiments, the chiral stationary phase is a member of theamylosic or cellulosic class of polysaccharides that is selected fromcellulose phenyl carbamate derivatives, such as cellulosetris(3,5-dimethylphenyl)carbamate (available from Daicel ChemicalIndustries, Ltd. (Daicel) as “Chiralcel® OD” or “Chiralpak® IB”, whereinthe carbamate derivative is bonded to the cellulosic backbone);cellulose tribenzoate derivatives, such as cellulose tri4-methylbenzoate (available from Daicel as “Chiralcel® OJ”); cellulosetricinnamate (available from Daicel as “Chiralcel® OK”); amylase phenyland benzyl carbamate derivatives, such as amylose tris[(S)-α-methylbenzylcarbamate] (available from Daicel as “Chiralpak® AS”); amylosetris(3,5-dimethylphenyl)carbamate (available from Daicel as “Chiralpak®AD” or “Chiralpak® IA”, wherein the carbamate derivative is bonded tothe amylosic backbone); amylose 3,4-substituted phenyl carbamate oramylose 4-substituted phenyl-carbamate; and amylose tricinnamate. Insome embodiments, the chiral stationary phase comprises Chiralpak® IA orChiralpak AD. In some embodiments, the chiral stationary phase comprisesChiralcel® OD. In some embodiments, the chiral stationary phase is amember of the Pirkle-phases family such as 3,5-dinitrobenzoylderivatives of phenylglycine (available from Regis Technologies Inc as“phenylglycine”; 5-dinitrobenzoyl derivative of leucine (available fromRegis Technologies Inc as“Leucine”);N-3,5-dinitrobenzoyl-3-amino-3-phenyl-2-(1,1-dimethylethyl)-propanoate(available from Regis Technologies Inc as “β-GEM 1”); dimethylN-3,5-dinitrobenzoyl-amino-2,2-dimethyl-4-pentenyl phosphonate(available from Regis Technologies Inc as “α-BURKE 2”);3-(3,5-dinitrobenzamido)-4-phenyl-β-lactam (available from RegisTechnologies Inc as “PIRKLE 1-J”); 3,5-Dintrobenzoyl derivative ofdiphenylethylenediamine (available from Regis Technologies Inc as“ULMO”); 4-(3,5-dinitro benzamido)tetrahydrophenanthrene (available fromRegis technologies Inc. as “(S,S) Whelk-O® 1” and “(R,R) Whelk-O® 1” or“(S,S) Whelk-O®2” and “(R,R) Whelk-O®2”); 3,5-dinitro-benzoyl derivativeof 1,2-diaminocyclohexane, (available from Regis technologies Inc. as“DACH-DNB). In some embodiments, the chiral stationary phase comprises“(S,S) Whelk-O® 1” or “(R,R) Whelk-O® 1.

In some embodiments, the particle diameter of the chiral stationaryphase is usually 1 to 300 μm, 2 to 100 μm, 5 to 75 μm, or 10 to 30 μm.

In some embodiments, the mobile phase is non-polar, polar protic oraprotic solvents or mixture thereof. In some embodiments, the mobilephase is a mixture of carbon dioxide and polar protic solvents. Suitablenon polar solvents include, for example, hydrocarbons, for instance,n-pentane, n-hexane, hexanes, n-heptane, heptanes, cyclohexane, andmethylcyclohexane. Suitable protic or aprotic solvents include, forexample, alcohols, in particular methanol, ethanol, 1-propanol,2-propanol, 1-butanol, 2-butanol, isobutanol, tert butanol, ethers, forinstance methyl tert butyl ether, esters, for instance ethylacetate,halogenated hydrocarbons and acetonitrile. In some embodiments, thenon-polar solvent is n-heptane. In some embodiments, the protic oraprotic solvent is ethanol, 2-propanol or methyl-tert-butyl ether. Insome embodiments, the mobile phase is a mixture of heptane and ethanol.In some embodiments, the ethanol is present in the mobile phase in anamount of about 10% to about 100%, about 10% to about 25%, or about 15%.In some embodiments, the mobile phase is a mixture of heptane and2-propanol. In some embodiments, the 2-propanol is present in the mobilephase in an amount of about 10% to about 100%, about 10% to about 25%,or about 20%. In some embodiments, the mobile phase is a mixture ofheptane and methyl-tert-butyl ether. In some embodiments, themethyl-tert-butyl ether is present in the mobile phase in an amount ofabout 10% to about 100%, about 75% to about 100%, or about 90% to about100%.

In some embodiments, the chromatography is carried out at a temperaturerange of about 0° C. to 50° C., about 10° C. to 30° C., or about 25° C.

In some embodiments, the desired enantiomer is recovered at anenantiomeric purity greater than about 90%, greater than about 98%, orgreater than about 99.0%. In some embodiments, the desired enantiomer isrecovered with a yield greater than about 70%, greater than about 90%,or greater than about 95%.

In some embodiments, the desired enantiomer is produced at a ratethroughput greater than about 0.1 kg, 0.4 kg, or 0.8 kg pure enantiomerper day per kilogram of stationary phase.

In some embodiments, the separated enantiomers are recovered afterevaporation under reduced pressure as concentrated oils.

In some embodiments, the mobile phase used in the chiral chromatographyprocess is recycled.

In some embodiments, the undesired enantiomer is racemized and reused asracemic feed for the chiral separation.

In some embodiments, the compound of Formula IX has the formula:

In some embodiments, the enantiomeric excess is equal to or greater thanabout 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%,about 99%, about 99.1%, about 99.2%, about 99.3%, about 99.4%, about99.5%, about 99.6%, about 99.7%, about 99.8%, about 99.9%, or about99.99%.

In some embodiments, the desired enantiomer is recovered in at least a91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% yield, and preferablygreater than 90% or 95% yield.

In some embodiments, the process further comprises reacting the compoundof Formula IX:

with a compound of Formula XI:

in the presence of a palladium catalyst, base, and a solvent underconditions and for a time sufficient to form a composition comprising anenantiomeric excess of the (R)- or (S)-enantiomer of a compound ofFormula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R_(c) and R_(d) are each independently C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups; and

X₂ is a tosylate group, a triflate group, iodo, chloro, or bromo.

In some embodiments, the process further comprises reacting the compoundof Formula IX:

with a compound of Formula X:

in the presence of a palladium catalyst, base, and a solvent underconditions and for a time sufficient to form a composition comprising anenantiomeric excess of the (R)- or (S)-enantiomer of a compound ofFormula Ia:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;

R_(c) and R_(d) are each independently C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups;

X₂ is a tosylate group, a triflate group, iodo, chloro, or bromo; and

P₁ is a protecting group.

In some embodiments, X₂ is bromo, iodo, or chloro. In some embodiments,X₂ is chloro.

The Suzuki coupling reactions can be initiated using a number ofpalladium(0) and palladium(II) catalysts and performed under conditionsknown in the art (see, e.g., Miyaura and Suzuki, Chem. Rev. 1995, 95,2457-2483, which is hereby incorporated in its entirety). In someembodiments, the palladium catalyst is Pd(PPh₃)₄ and Pd(dppf)₂Cl₂.

In some embodiments, the palladium catalyst istetrakis(triphenylphosphine)palladium(0) ortetrakis(tri(o-tolyl)phosphine)palladium(0). In some embodiments, thepalladium catalyst is tetrakis(triphenylphosphine)palladium(0).

In some embodiments, the palladium catalyst loading is from about 1×10⁻⁴to about 0.1 equivalents. In some embodiments, the palladium catalystloading is from about 0.0010 to about 0.0015 equivalents. In someembodiments, the stoichiometric ratio of the compound of Formula X or XIto the compound of Formula IX is from about 1:1.05 to about 1:1.35.

In some embodiments, the solvent comprises water and an organic solvent.In some embodiments, the organic solvent is 1,4-dioxane, 1-butanol,1,2-dimethoxyethane (DME), 2-propanol, toluene or ethanol, or acombination thereof. In some embodiments, the organic solvent comprisesDME. In some embodiments, the organic solvent comprises DMF.

In some embodiments, the base is an inorganic base. In some embodiments,the base is an organic base. In some embodiments, the base is an alkalimetal carbonate. In some embodiments, the base is potassium carbonate(K₂CO₃). In some embodiments, two to five equivalents of base (e.g.,K₂CO₃) are used.

In some embodiments, the Suzuki coupling reaction is conducted at atemperature of about 80 to about 100° C. In some embodiments, thereaction is carried out for two to twelve hours. In some embodiments,the compound of Formula Ia or III can be optionally isolated fromaqueous work-up of the Suzuki coupling reaction mixture or directlyused.

Appropriate P₁ groups and deprotection conditions are provided supra.

In some embodiments, the present invention provides a process ofpreparing a racemate of a compounds of Formula IX

comprising reacting a compound of Formula XV:

with a compound of Formula D-1:

in the presence of a base to produce compound of Formula IX;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

R_(c) and R_(d) are each independently C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom to which the oxygen atoms are attached, forma 5- to 6-membered heterocyclic ring, which is optionally substitutedwith 1, 2, 3, or 4 C₁₋₄ alkyl groups.

In some embodiments, the aza-Michael addition is conducted in an organicsolvent at room temperature in the presence of a catalytic amound ofbase. The base can be suitable solvent or base for aza-Michaelreactions. In some embodiments, the solvent is acetonitrile ordimethylformide (DMF). In some embodiments, the base is atetraalkylammonium halide, tetraalkylammonium hydroxide, guanidine,amidine, hydroxide, alkoxide, silicate, alkali metal phosphate, oxide,tertiary amine, alkali metal carbonate, alkali metal bicarbonate, alkalimetal hydrogen phosphate, phosphine, or alkali metal salt of acarboxylic acid. In some embodiments, the Michael addition catalyst istetramethyl guanidine, 1,8-diazabicyclo(5.4.0)undec-7-ene,1,5-diazabicyclo(4.3.0)non-5-ene, 1,4-diazabicyclo(2.2.2)octane,tert-butyl ammonium hydroxide, sodium hydroxide, potassium hydroxide,sodium methoxide, sodium ethoxide, tripotassium phosphate, sodiumsilicate, calcium oxide, triethylamine, sodium carbonate, potassiumcarbonate, sodium bicarbonate, potassium bicarbonate, potassium hydrogenphosphate, triphenyl phosphine, triethyl phosphine, potassium acetate,or potassium acrylate. In some embodiments, the base is1,8-diazabicyclo[5.4.0]unde-7-ene (DBU) or potassium carbonate. In someembodiments, the base is DBU. In some embodiments, the base is presentin a catalytic amount. In some embodiments, the amount of base is about0.1 to about 5 equivalents, or about 0.5 to about 3 equivalents. In someembodiments, the reaction is complete in about 10 to about 24 hours.

In some embodiments, the process further comprises treating the compoundof Formula Ia under deprotection conditions sufficient to provide acomposition comprising an enantiomeric excess of a (R)- or(S)-enantiomer of a compound of Formula III:

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

Appropriate P₁ groups and deprotection methods include, but are notlimited to those described supra.

In some embodiments, the process further comprises reacting a compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

IV. Chiral Enrichment of Racemates of Formula Ia and Racemization ofUndesired Enantiomers of Formula Ia

Racemates of Formula Ia can be formed by a Michael addition process inScheme 6 below. Accordingly, a compound of Formula IV can be reactedwith an acrylonitrile of Formula D-1 to form a racemate of Formula Ia.The racemate of Formula Ia can then be separated by chiral columnchromatography to give a composition comprising an enantiomeric excessof the (R)- or (S)-enantiomer of the compound of Formula Ia. Theprotecting group can then be removed to produce an enantiomeric excessof the (R)- or (S)-enantiomer of the compound of Formula III.

Accordingly, in some embodiments, the present invention provides aprocess of preparing a composition comprising an enantiomeric excess ofthe (R)- or (S)-enantiomer of a compound of Formula Ia:

comprising passing a composition comprising a racemate of a compound ofFormula Ia through a chiral chromatography unit using a mobile phase andcollecting a composition comprising an enantiomeric excess of the (R)-or (S)-enantiomer of a compound of Formula Ia;wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl;and

P₁ is a protecting group.

In some embodiments, the chiral chromatography unit is a preparativehigh performance liquid chromatography (HPLC) system equipped with achiral column, which is packed with a chiral stationary phase. In someembodiments, the chiral column is Chiralpak® IA. In some embodiments,the chiral column is ChiralCel® OD-H. In some embodiments, thechromatography unit is stimulated moving bed (SMB) chromatography unitequipped with a set of eight columns, each packed with a chiralstationary phase. In some embodiments, the chiral stationary phase iscellulose modified chiral stationary phase (CSP, Chiral Technologies Insome embodiments, the mobile phase comprises ethanol and hexanes. Insome embodiments, the mobile phase comprises about 1:9 ratio of ethanolto hexanes by volume. In some embodiments, the mobile phase comprisesabout 15% ethanol and about 85% hexanes by volume. In some embodiments,the mobile phase comprise ethanol and hexanes, wherein the ethanol ispresent in an amount of about 25% to about 10% by volume. In someembodiments, the mobile phase comprises isopropanol and hexanes, whereinthe isopropanol is present in an amount of about 25% to about 10% byvolume. In some embodiments, the chromatography unit is kept at roomtemperature. In some embodiments, the mobile phase is passed at a flowrate of about 1 mL per minute to about 20 mL per minute. In someembodiments, the mobile phase is passed at a flow rate of about 1 mL perminute. In some embodiments, the mobile phase is passed at a flow rateof about 18 mL per minute. In some embodiments, the eluent is monitoredby ultraviolet (UV) spectroscopy. In some embodiments, the eluent ismonitored by ultraviolet spectroscopy at about 220 nm. Collection of theportion of the eluent containing the enantiomerically enrichedcomposition can be determined by detection of the elution of the desiredenantiomer by UV spectroscopy. Determination of the % ee (enantiomericexcess) of the composition can then be determined by analytical chiralHPLC.

In some embodiments, the enantiomeric excess is equal to or greater thanabout 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%,about 99%, about 99.1%, about 99.2%, about 99.3%, about 99.4%, about99.5%, about 99.6%, about 99.7%, about 99.8%, about 99.9%, or about99.99%.

In some embodiments, the chiral chromatography is performed using apreparative high performance liquid chromatography (HPLC) systemequipped with a chromatographic column, which is packed with a chiralstationary phase. In some embodiments, the column is packed with chiralstationary phase made of amylose tris(3,5-dimethylphenyl carbamate)immobilized on silica gel (available from Daicel as “Chiralpak® IA”). Insome embodiments, the column is packed with a chiral stationary phasemade of cellulose tris(3,5-dimethylphenyl carbamate) coated on silicagel (available from Daicel as ‘Chiralcel® OD”). In some embodiments, thechromatography process is a continuous chromatography process such assimulated moving bed (SMB) chromatography or Varicol process using aunit equipped with a set of 3 to 12 columns, preferably 5 to 10, mostpreferably 5 to 8, each column packed with the same chiral stationaryphase. In some embodiments, the column is packed with chiral stationaryphase made of amylose tris(3,5-dimethylphenyl carbamate) immobilized onsilica gel (available from Daicel as “Chiralpak® IA”). In someembodiments, the column is packed with a chiral stationary phase made ofcellulose tris(3,5-dimethylphenyl carbamate) coated on silica gel(available from Daicel as “Chiralcel® OD”). In some embodiments, thechiral stationary phase is a silica gel-based stationary phase coatedwith 4-(3,5-dinitro benzamido)tetrahydrophenanthrene (available fromRegis Technologies as “(S,S) Whelk-O®1”). In some embodiments, themobile phase comprises ethanol and hexanes. In some embodiments, thehexanes are replaced by heptanes, n-heptane, cyclohexane ormethylcyclohexane. In some embodiments, the ethanol is present in anamount of about 10% to about 100% by volume, about 10% to about 25% byvolume, or about 15% by volume. In some embodiments, the mobile phasecomprises isopropanol and hexanes. In some embodiments the hexanes arereplaced by heptanes, n-heptane, cyclohexane or methylcyclohexane. Insome embodiments, the isopropanol is present in an amount of about 10%to about 25% by volume. In some embodiments, the mobile phase comprisesmethyl-tert-butyl ether and hexanes. In some embodiments, the hexanesare replaced by heptanes, n-heptane, cyclohexane or methylcyclohexane.In some embodiments, the methyl-tert-butyl ether is present in an amountof about 10% to about 100% by volume, about 50% to about 100% by volume,or about 90% to about 100% by volume. In some embodiments, the mobilephase comprises ethyl acetate and hexanes. In some embodiments, thehexanes are replaced by heptanes, n-heptane, cyclohexane ormethylcyclohexane. In some embodiments, the ethyl acetate is present inamount of about 10% to about 100% by volume, about 50% to about 100% byvolume, or about 75% by volume. In some embodiments, the mobile phasecomprises tetrahydrofuran and hexanes. In some embodiments, the hexanesare replaced by heptanes, n-heptane, cyclohexane or methylcyclohexane.In some embodiments, the tetrahydrofuran is present in an amount ofabout 10% to about 100% by volume, about 10% to about 50% by volume, orabout 25% by volume. In some embodiments, the chromatography unit isoperated at a temperature of about 5° C. to about 50° C., at about 10°C. to about 30° C., or at about 25° C., or at ambient temperature.

In some embodiments, the chromatographic method employed is batchpreparative chromatography, supercritical fluide chromatography (SFC), acyclojet process, a continuous multicolumn chromatography process, asimulated moving bed process, a Varicol™ process, or a PowerFeedprocess.

In some embodiments, the chiral stationary phase comprises aninteracting agent which is an enantiomerically enriched resolving agent,immobilized to an inert carrier material by, for example, chemicallybinding or by insolubilizing via cross-linking. The suitable inertcarrier material can be macroporous, e.g crosslinked polystyrene,polyacrylamide, polyacrylate, alumina, kieselgur (diatomaceous), quartz,kaolin, magnesium oxide, titanium dioxide or silica gel. In someembodiments, the inert carrier material is Silicagel.

In some embodiments, the chiral stationary phase is a member of theamylosic or cellulosic class of polysaccharides that is selected fromcellulose phenyl carbamate derivatives, such as cellulosetris(3,5-dimethylphenyl)carbamate (available from Daicel ChemicalIndustries, Ltd. (Daicel) as “Chiralcel® OD” or “Chiralpak® IB”, whereinthe carbamate derivative is bonded to the cellulosic backbone);cellulose tribenzoate derivatives, such as cellulose tri4-methylbenzoate (available from Daicel as “Chiralcel® OJ”); cellulosetricinnamate (available from Daicel as “Chiralcel® OK”); amylase phenyland benzyl carbamate derivatives, such as amylose tris[(S)-α-methylbenzylcarbamate] (available from Daicel as “Chiralpak® AS”); amylosetris(3,5-dimethylphenyl)carbamate (available from Daicel as “Chiralpak®AD” or “Chiralpak® IA”, wherein the carbamate derivative is bonded tothe amylosic backbone); amylose 3,4-substituted phenyl carbamate oramylose 4-substituted phenyl-carbamate; and amylose tricinnamate. Insome embodiments, the chiral stationary phase comprises Chiralpak® IA orChiralpak AD. In some embodiments, the chiral stationary phase comprisesChiralcel® OD. In some embodiments, the chiral stationary phase is amember of the Pirkle-phases family such as 3,5-dinitrobenzoylderivatives of phenylglycine (available from Regis Technologies Inc as“phenylglycine”; 5-dinitrobenzoyl derivative of leucine (available fromRegis Technologies Inc as“Leucine”);N-3,5-dinitrobenzoyl-3-amino-3-phenyl-2-(1,1-dimethylethyl)-propanoate(available from Regis Technologies Inc as “β-GEM 1”); dimethylN-3,5-dinitrobenzoyl-amino-2,2-dimethyl-4-pentenyl phosphonate(available from Regis Technologies Inc as “α-BURKE 2”);3-(3,5-dinitrobenzamido)-4-phenyl-β-lactam (available from RegisTechnologies Inc as “PIRKLE 1-J”); 3,5-Dintrobenzoyl derivative ofdiphenylethylenediamine (available from Regis Technologies Inc as“ULMO”); 4-(3,5-dinitro benzamido)tetrahydrophenanthrene (available fromRegis technologies Inc. as “(S,S) Whelk-O® 1” and “(R,R) Whelk-O® 1” or“(S,S) Whelk-O®2” and “(R,R) Whelk-O®2”); 3,5-dinitro-benzoyl derivativeof 1,2-diaminocyclohexane, (available from Regis technologies Inc. as“DACH-DNB). In some embodiments, the chiral stationary phase comprises“(5,5) Whelk-O® 1” or “(R,R) Whelk-O® 1.

In some embodiments, the particle diameter of the chiral stationaryphase is usually 1 to 300 μm, 2 to 100 μm, 5 to 75 μm, or 10 to 30 μm.

In some embodiments, the mobile phase is non-polar, polar protic oraprotic solvents or mixture thereof. In some embodiments, the mobilephase is a mixture of carbon dioxide and polar protic solvents. Suitablenon polar solvents include, for example, hydrocarbons, for instance,n-pentane, n-hexane, hexanes, n-heptane, heptanes, cyclohexane, andmethylcyclohexane. Suitable protic or aprotic solvents include, forexample, alcohols, in particular methanol, ethanol, 1-propanol,2-propanol, 1-butanol, 2-butanol, isobutanol, tert butanol, ethers, forinstance methyl tert butyl ether, esters, for instance ethylacetate,halogenated hydrocarbons and acetonitrile. In some embodiments, thenon-polar solvent is n-heptane. In some embodiments, the protic oraprotic solvent is ethanol, 2-propanol or methyl-tert-butyl ether. Insome embodiments, the mobile phase is a mixture of heptane and ethanol.In some embodiments, the ethanol is present in the mobile phase in anamount of about 10% to about 100%, about 10% to about 25%, or about 15%.In some embodiments, the mobile phase is a mixture of heptane and2-propanol. In some embodiments, the 2-propanol is present in the mobilephase in an amount of about 10% to about 100%, about 10% to about 25%,or about 20%. In some embodiments, the mobile phase is a mixture ofheptane and methyl-tert-butyl ether. In some embodiments, themethyl-tert-butyl ether is present in the mobile phase in an amount ofabout 10% to about 100%, about 75% to about 100%, or about 90% to about100%.

In some embodiments, the chromatography is carried out at a temperaturerange of about 0° C. to 50° C., about 10° C. to 30° C., or about 25° C.

In some embodiments, the desired enantiomer is recovered at anenantiomeric purity greater than about 90%, greater than about 98%, orgreater than about 99.0%. In some embodiments, the desired enantiomer isrecovered with a yield greater than about 70%, greater than about 90%,or greater than about 95%.

In some embodiments, the desired enantiomer is produced at a ratethroughput greater than about 0.1 kg, 0.4 kg, or 0.8 kg pure enantiomerper day per kilogram of stationary phase.

In some embodiments, the separated enantiomers are recovered afterevaporation under reduced pressure as concentrated oils.

In some embodiments, the mobile phase used in the chiral chromatographyprocess is recycled.

In some embodiments, the undesired enantiomer is racemized and reused asracemic feed for the chiral separation.

In some embodiments, the desired enantiomer is recovered in at least a91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% yield, or preferablygreater than 90% or 95% yield.

Alternatively, the racemate of Formula Ia can be reacted with a chiralacid (E-1), such as (+)-dibenzoyl-D-tartaric acid, to form a chiral salt(E-2) (Scheme 7). After crystallization, filtration, and treatment withbase, a composition comprising an enantiomeric excess of the (R)- or(S)-enantiomer of the compound of Formula Ia is produced. The protectinggroup can then be removed to produce an enantiomeric excess of the (R)-or (S)-enantiomer of the compound of Formula III.

Accordingly, in some embodiments, the present invention provides aprocess of preparing a composition comprising an enantiomeric excess ofa (R)- or (S)-enantiomer of a compound of Formula Ia:

comprising:

(a) reacting a composition comprising a racemate of a compound ofFormula Ia with a chiral acid in the presence of a solvent to form asalt of a compound of Formula Ia;

(b) separating a composition comprising an enantiomer excess of a chiralsalt of the (R)- or (S)-enantiomer of the compound of Formula Ia; and

(c) treating the chiral salt with a base to form a compositioncomprising an enantiomeric excess of the (R)- or (S)-enantiomer of thecompound of Formula Ia;

wherein:

* indicates a chiral carbon;

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆-fluoroalkyl;and

P₁ is a protecting group.

Any chiral acid useful for chiral resolution can be used. In someembodiments, the chiral acid is selected from optically active forms ofmandelic acid, 2-chloromandelic acid, camphorsulfonic acid, tartaricacid, lactic acid, malic acid, 3-bromocamphor-8-sulfonic acid,3-bromocamphor-10-sulfonic acid, 10-camphorsulfonic acid, dibenzoyltartaric acid, di-p-toluoyltartaric acid,2-amino-7,7-dimethylbicyclop[2,2,1]heptan-1-methylene sulfonic acid, and2-acrylamide-7,7-dimethylbicyclo[2,2,1]heptan-1-methylene sulfonic acid.In some embodiments, the chiral acid is (+)-dibenzoyl-D-tartaric acid.

In some embodiments, the solvent comprises acetonitrile,tetrahydrofuran, acetone, or combination thereof. In some embodiments,the solvent is about a 90:15:15 ratio by volume of acetonitrile,tetrahydrofuran, and acetone (15.0 mL, 0.204 mol).

In some embodiments, the enantiomeric excess is equal to or greater thanabout 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%,about 99%, about 99.1%, about 99.2%, about 99.3%, about 99.4%, about99.5%, about 99.6%, about 99.7%, about 99.8%, about 99.9%, or about99.99%.

In some embodiments, the separating involves cooling the solvent toprecipitate the chiral salt. In some embodiments, the separatinginvolves adding a second solvent to precipitate the chiral salt. In someembodiments, the separating comprises filtering the solvent to recoverthe chiral salt. In some embodiments, the solvent comprisesacetonitrile, tetrahydrofuran, acetone, or combination thereof. In someembodiments, the reacting is conducted at a temperature of about roomtemperature to about 60° C.

Any base suitable for preparing the free base of the chiral salt can beutilized in the process. In some embodiments, the base is an alkalimetal or alkaline earth metal hydroxide or carbonate. In someembodiments, the base is an alkali metal hydroxide. In some embodiments,the base is sodium hydroxide. In some embodiments, the treatingcomprises adding an aqueous solution of base to a solution of the chiralsalt, followed by separation of the solution from the aqueous solution.In some embodiments, the process further comprises removal of thesolvent.

In addition to the processes for chiral enrichment described supra,undesired enantiomers of compounds of Formula Ia can be converted toracemic material by base-catalyzed retro-Michael addition to form thecompound of Formula IV, followed by reaction with the acrylonitrile ofFormula D-1 to produce the racemic Michael adduct of Formula Ia as shownin Scheme 8. Alternatively, the undesired enantiomer of Formula Ia canbe epimerized in the presence of a Michael acceptor of Formula D-1 togive the racemate of Formula Ia as shown in Scheme 8. The racemate canthen be resolved to give the desired enantiomer by the chiral columnseparation and chiral salt methods described supra.

Accordingly, the present invention provides a process of preparing acomposition comprising a racemate of a compound of Formula Ia:

comprising:

a) treating a composition comprising an enantiomeric excess of the (R)-or (S)-enantiomer of a compound of Formula Ia with a compound of FormulaD-1

in the presence of a first base under conditions sufficient to form acompound of Formula IV:

and

(b) reacting a compound of Formula IV with a compound of Formula D-1 inthe presence of a second base;

wherein:

* indicates a chiral carbon;

P₁ is a protecting group; and

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl.

In some embodiments, the first base is an alkali metal or alkaline earthmetal base. In some embodiments, the first base is an alkali metal oralkaline earth metal base alkoxide, hydroxide or carbonate. In someembodiments, the first base is an alkali metal or alkaline earthcarbonate. In some embodiments, the first base is an alkaline earthcarbonate. In some embodiments, the first base is cesium carbonate. Insome embodiments, the first base is an alkali metal t-butoxide. In someembodiments, the first base is potassium t-butoxide.

In some embodiments, the second step is conducted in an organic solventat room temperature in the presence of a catalytic amound of the secondbase. The second base can be suitable solvent or second base foraza-Michael reactions. In some embodiments, the solvent is acetonitrileor dimethylformide (DMF). In some embodiments, the second base is atetraalkylammonium halide, tetraalkylammonium hydroxide, guanidine,amidine, hydroxide, alkoxide, silicate, alkali metal phosphate, oxide,tertiary amine, alkali metal carbonate, alkali metal bicarbonate, alkalimetal hydrogen phosphate, phosphine, or alkali metal salt of acarboxylic acid. In some embodiments, the base is tetramethyl guanidine,1,8-diazabicyclo(5.4.0)undec-7-ene, 1,5-diazabicyclo(4.3.0)non-5-ene,1,4-diazabicyclo(2.2.2)octane, tert-butyl ammonium hydroxide, sodiumhydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide,tripotassium phosphate, sodium silicate, calcium oxide, triethylamine,sodium carbonate, potassium carbonate, sodium bicarbonate, potassiumbicarbonate, potassium hydrogen phosphate, triphenyl phosphine, triethylphosphine, potassium acetate, or potassium acrylate. In someembodiments, the second base is 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU)or potassium carbonate. In some embodiments, the second base is DBU. Insome embodiments, the base is present in a catalytic amount. In someembodiments, the amount of second base is about 0.1 to about 5equivalents, or about 0.5 to about 3 equivalents, or about 0.1 to about0.5 equivalents. In some embodiments, the reaction is complete in about1 to about 3 hours.

Alternatively, the present invention further provides a process ofpreparing a composition comprising a racemate of a compound of FormulaIa:

comprising treating a composition comprising an enantiomeric excess ofthe (R)- or (S)-enantiomer of a compound of Formula Ia with a compoundof Formula D-1:

in the presence of a base under conditions sufficient to form theracemate of the compound of Formula Ia;wherein:

* indicates a chiral carbon;

P₁ is a protecting group; and

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl.

In some embodiments, the base is an alkali metal or alkaline earth metalbase. In some embodiments, the base is an alkali metal or alkaline earthmetal base alkoxide, hydroxide or carbonate. In some embodiments, thebase is an alkali metal or alkaline earth carbonate. In someembodiments, the base is an alkaline earth carbonate. In someembodiments, the base is cesium carbonate. In some embodiments, the baseis an alkali metal t-butoxide. In some embodiments, the base ispotassium t-butoxide.

The racemate of compounds of Formula Ia:

can be prepared by a process comprising treating a compound of FormulaIV:

with a compound of Formula D-1:

under conditions sufficient to form the racemate of the compound ofFormula Ia;wherein:

* indicates a chiral carbon;

P₁ is a protecting group; and

R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆ alkyl, and C₁₋₆ fluoroalkyl.

3-Substituted acrylnitriles of Formula D-1 are prepared as shown inScheme 9. Olefination of an aldehyde of Formula D-2, such ascyclopentanecarbaldehyde or cyclopropanecarbaldehyde, with a Wittig-typereagent having a ylide of formula —CH₂CN, such as diethylcyanomethylphosphonate, is conducted in an organic solvent, such as THF,under the influence a base, such as potassium tert-butoxide, at about 0to about 5° C. In some embodiments, the resulting 3-substitutedacrylnitriles of Formula D-1 can be purified by vacuum distillation.

Accordingly, in some embodiments, the compound of Formula D-1:

is prepared by a process comprising reacting a compound of Formula D-2:

with a Wittig-type reagent having a ylide of formula —CH₂CN in thepresence of a base; wherein R₁ is selected from C₃₋₇ cycloalkyl, C₁₋₆alkyl, and C₁₋₆ fluoroalkyl.

As used herein, the term “Wittig-type reagent” refers to reagents usedin the Wittig reaction, the Wadsworth-Emmons reaction, and theHorner-Wittig reaction as described in the art (see e.g., Carey andSundberg, Advanced Organic Chemistry, Part B: Reactions and Synthesis,4th ed., Kluwer Academic/Plenum Publishers:New York, pages 111-119(2001); and March, Advanced Organic Chemistry: Reactions, Mechanisms,and Structure, 3rd ed., John Wiley & Sons:New York, pages 845-855(1985), each of which is incorporated herein by references in itsentirety). Exemplative Wittig-type reagents containing a cyanomethyl orcyanomethyl glide group include, but are not limited to, compounds ofgeneral formula (R′O)₂P(═O)-L-R¹, R″₃P(+)-L(−)-R¹, R″₃P(+)-L-R¹X;R″₂P(═O)-L-R¹, and (R′N)₂P(═O)-L-R¹, wherein R′ is C₁₋₆ alkoxy oroptionally substituted phenyl; R″ is optionally substituted phenyl; L is—CH₂— or —CH—; and R¹ is cynao; and X is an anion (e.g., halo anion,such as chloride). In some embodiments, the Wittig-type reagent isdiethyl cyanomethyl phosphate. In some embodiments, the reacting of thecompound of Formula D-2 with the Wittig-type reagent in the presence ofa base. In some embodiments, the base is a strong base. In someembodiments, the base is potassium t-butoxide, sodium t-butoxide, sodiumhydride, sodium ethoxide, sodium hydroxide, potassium carbonate, orsodium carbonate. In some embodiments, the base is an alkali metalalkoxide. In some embodiments, the base is an alkali metal t-butoxide.In some embodiments, the base is potassium t-butoxide. In someembodiments, the olefination of the aldehyde of Formula D-2 with aWittig-type reagent is conducted in an organic solvent, such as THF,under the influence a base, such as potassium tert-butoxide, at atemperature from about 0 to about 5° C. In some embodiments, the base ispresent in about 1 to about 1.2 equivalents, or about 1.05 to about 1.1equivalents, with respect to the compound of Formula D-2. In someembodiments, the Wittig-type reagent is present in about 1 to about 1.2equivalents, or about 1.05 to about 1.1 equivalents with respect to thecompound of Formula D-2. In some embodiments, the Wittig-type reagent is(methoxymethyl)triphenylphosphinium chloride.

In other embodiments, the processes further comprise reacting thecompound of Formula Ia under deprotection conditions to form a compoundof Formula III:

Appropriate P₁ groups and deprotection methods include, but are notlimited to those described supra.

In some embodiments, the process further comprises reacting a compoundof Formula III with phosphoric acid to form a phosphate salt of thecompound of Formula III.

V. Routes to Intermediate Compounds

i) Higher Yield Routes to Intermediate Compounds of Formula IV

Compounds of Formula IV are important intermediates in the varioussynthetic routes for the compounds of Formula III described supra. Thesecompounds are generally formed by Suzuki coupling processes. Suzukicoupling of protected 7H-pyrrolo[2,3-d]pyrimidine derivative of FormulaX with a unprotected pyrazole borate derivative of Formula XV using apalladium catalyst result in a lower yield (Scheme 10). Without wishingto be bound by any particular theory, it is believed that the loweryields result from interference of unprotected amine functionality inthe Suzuki coupling reaction.

Accordingly, a new process for preparing the compound of Formula IV wasdeveloped involving the use of protected pyrazole borate derivative ofFormula XIII (Scheme 11). Accordingly, the compound of Formula XIII canbe generated and then reacted with the protected7H-pyrrolo[2,3-d]pyrimidine derivative of Formula X to form a compoundof Formula XII, followed by deprotection to give the compound of FormulaIV. In some embodiments, the compound of Formula XIII can be formed byin-situ protection of pyrazole pinacol borate. For example, when P₂ is1-(ethoxy)ethyl, a pyrazol-4-yl pinacol borate can be reacted with vinylether in-situ to generate the protected compound of Formula XIII. TheSuzuki coupling reaction between the protected pyrazole pinacol borateof Formula XIII and the compound of Formula X then proceeds smoothlyunder the typical Suzuki reaction conditions to generate the compound ofFormula IV in higher yield after the acidic work-up of the correspondingcoupling intermediate of Formula XII.

In other embodiments, the compound of Formula XIII is the isolated andfully characterized compound. For example, the use of isolated, fullycharacterized compound of Formula XIII, wherein P₂ is 1-(ethoxy)ethyland the borate moiety is a pinacol group, afforded the product ofFormula XII, and subsequently the compound of Formula IV in better yieldand purity.

In other embodiments, the compound of Formula X can be in-situ generatedfrom a compound of Formula XI and then subsequently reacted with thecompound of Formula XIII. This eliminates the necessity of having toisolate and purify the compound of Formula X during large-scaleproduction. For example, when P₁ is SEM, the compound of Formula XI canbe reacted with sodium hydride and SEM chloride to generate the compoundof Formula X in situ (Scheme 12).

Accordingly, the present invention provides a process of preparing acompound of Formula XII:

comprising reacting a compound of Formula X:

with a compound of Formula XIII:

in the presence of a palladium catalyst, base, and a solvent, to form acompound of Formula XII;wherein:

X₂ is a tosylate group, a triflate group, iodo, chloro, or bromo;

P₁ and P₂ are each independently a protecting group;

R_(c) and R_(d) are each independently H or C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom, form a 5- to 6-membered heterocyclic ring,which is optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups.

In some embodiments, the process further comprises a process forpreparing a compound of Formula IV:

comprising reacting the compound of Formula XII under deprotectionconditions to produce a compound of Formula IV;wherein:

P₁ and P₂ are each independently a protecting group; and

R_(c) and R_(d) are each independently H or C₁₋₆ alkyl; or

R_(c) and R_(d), together with the oxygen atoms to which they areattached and the boron atom, form a 5- to 6-membered heterocyclic ring,which is optionally substituted with 1, 2, 3, or 4 C₁₋₄ alkyl groups.

In some embodiments, the compound of Formula XIII is:

In some embodiments, X₂ is chloro, bromo, or iodo. In some embodiments,X₂ is chloro.

The Suzuki coupling reaction can be initiated using a number ofpalladium(0) and palladium(II) catalysts and performed under conditionsknown in the art (see, e.g., Miyaura and Suzuki, Chem. Rev. 1995, 95,2457-2483, which is hereby incorporated in its entirety). In someembodiments, the palladium catalyst is Pd(PPh₃)₄ and Pd(dppf)₂Cl₂.

In some embodiments, the palladium catalyst istetrakis(triphenylphosphine)palladium(0) ortetrakis(tri(o-tolyl)phosphine)palladium(0). In some embodiments, thepalladium catalyst is tetrakis(triphenylphosphine)palladium(0).

In some embodiments, the palladium catalyst loading is from about 1×10⁻⁴to about 0.1 equivalents. In some embodiments, the palladium catalystloading is from about 0.0010 to about 0.0015 equivalents. In someembodiments, the stoichiometric ratio of the compound of Formula X tothe compound of Formula XIII is from about 1:1.05 to about 1:1.35.

In some embodiments, the solvent comprises water and an organic solvent.In some embodiments, the organic solvent is 1,4-dioxane, 1-butanol,1,2-dimethoxyethane (DME), 2-propanol, toluene or ethanol, or acombination thereof. In some embodiments, the organic solvent comprisesDME. In some embodiments, the organic solvent comprises DMF.

In some embodiments, the base is an inorganic base. In some embodiments,the base is an organic base. In some embodiments, the base is an alkalimetal carbonate. In some embodiments, the base is potassium carbonate(K₂CO₃). In some embodiments, two to five equivalents of base (e.g.,K₂CO₃) are used.

In some embodiments, the Suzuki coupling reaction is conducted at atemperature of about 80 to about 100° C. In some embodiments, thereaction is carried out for two to twelve hours. In some embodiments,the compound of Formula XII can be optionally isolated from aqueouswork-up of the Suzuki coupling reaction mixture or directly used.

In some embodiments, the compound of X is selected from those in Scheme13 and can be formed starting from a compound of Formula XI as shown. Insome embodiments, X₂ is chloro. In some embodiments, the compounds ofFormula X are isolated or in-situ generated as the starting materialsfor subsequent Suzuki reaction with or without further purification. Insome embodiments, the P₁ protecting group is one of those listed supra.

Appropriate P₂ protecting groups include, but are not limited to theprotecting groups for amines delineated in Wuts and Greene, ProtectiveGroups in Organic Synthesis, 4th ed., John Wiley & Sons: New Jersey,pages 696-887 (and, in particular, pages 872-887) (2007), which isincorporated herein by reference in its entirety. In some embodiments,P₂ is a protecting group which can be selectively removed underconditions which do not displace the P₁ protecting group. In someembodiments, P₂ is protecting group which can be removed under acidicconditions at room temperature, at a temperature from about 15° C. toabout 40° C., or at a temperature from about 15° C. to about 30° C. Insome embodiments, P₂ is a group which is deprotected under roomtemperature acidic conditions. In some embodiments, P₂ is1-(ethoxy)ethyl, tri(C₁₋₆ alkyl)silyl (e.g., t-butyldimethylsilyl ortriisopropylsilyl), p-methoxybenzyl (PMB), triphenylmethyl (Tr),diphenylmethyl, hydroxymethyl, methoxymethyl (MOM), diethoxymethyl, ort-butyldimethylsilylmethyl. In some embodiments, P₂ is 1-(ethoxy)ethyl.

Treatment of the compound of Formula XII to remove the P₂ group can beaccomplished by methods known in the art for the removal of particularprotecting groups for amines, such as those in Wuts and Greene,Protective Groups in Organic Synthesis, 4th ed., John Wiley & Sons: NewJersey, pages 696-887 (and, in particular, pages 872-887) (2007), whichis incorporated herein by reference in its entirety. In someembodiments, the treating comprises treating the compound of Formula XIIunder acidic conditions (e.g., hydrochloric acid or trifluoroaceticacid) at room temperature, at a temperature from about 15° C. to about40° C., or at a temperature from about 15° C. to about 30° C. In someembodiments, the treating comprises treating the compound of Formula XIIwith an aqueous solution of from about 1 N to about 5 N hydrochloricacid at a temperature of from about 10° C. to about 30° C.

Appropriate P₁ groups include, but are not limited to, those describedsupra.

Compounds of Formula X can be formed by protecting a compound of FormulaXI. Accordingly, in some embodiments, the process for preparing acompound of Formula X, comprises treating a compound of Formula XI:

to add a protecting group in order to form a compound of Formula X

wherein:

X₂ is a tosylate group, a triflate group, iodo, chloro, or bromo; and

P₁ is a protecting group.

In some embodiments, the compound of Formula XI can be deprotonated witha base, preferably with sodium hydride (NaH), in an organic solvent,such as THF, 1,4-dioxane, 1,2-dimethoxyethane (DME), orN,N-dimethylacetamide (DMAC), at low temperature, preferably at atemperature of about 0 to about 5° C. before being treated with anelectrophile, such as chloromethyl pivalate (POM-Cl) ortrimethylsilylethoxymethyl chloride (SEM-Cl) to add the protectinggroup, P₁. The protected compound X is isolated or in-situ generated asthe starting material for subsequent Suzuki reaction with or withoutfurther purification.

The intermediates formed from the processes described herein can be usedas appropriate in the other processes described herein.

ii. Preparation of Pinacol Borates of Formula C-9

The present invention further provides methods of preparing pyrazolpinacol borates of Formula XVI, which are useful in the processesdescribed herein. A specific subset of the compounds of Formula XVI arethe 4-substituted pyrazole borate derivatives of Formula XIIIa, whichcan be substituted for the compounds of Formula XIII above.

The compounds of XVI can be produced by the methods shown in Scheme 14.First, pyrazole is reacted with a halogenating agent to give themonohalo or dihalo pyrazole of Formula XIX (wherein X₃ is iodo or bromoand m is 1 or 2). The compound of Formula XIX is then protected to givethe protected monohalo or dihalo pyrazole of Formula XVIII. The compoundof Formula XVIII can then be treated with an alkyl Grignard oralkyllithium reagent, followed by treatment with a2-alkoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane reagent of FormulaXVII, to form the desired pinacol borate of Formula XVI. In someembodiments, the P₃ protecting group is one which is stable to anaqueous workup of the Grignard or lithium reaction (e.g., wherein P₃ is1-(ethoxy)ethyl). In other cases, P₃ is a protecting group is not stableto the aqueous workup of the Grignard or alkyllithium reaction. In thiscase, an additional protecting step will be needed to add the protectinggroup, P₃. In some embodiments, P₃ will be selected from the groupslisted supra for P₂, for ease of processing.

Accordingly, in some embodiments, the present invention provides aprocess for preparing a compound of Formula XVI:

comprising:

(a) reacting a compound of Formula XVIII

with about 1 or more equivalents of an C₁₋₆ alkyl Grignard reagent orC₁₋₆ alkyl lithium reagent followed by treating with about 1 or moreequivalents of a compound of Formula XVII:

and

(b) optionally, reprotecting the product of step (a) to give a compoundof Formula XVI;

wherein:

P₃ is a protecting group;

X₃ is halogen;

R₄ is C₁₋₆ alkyl; and

m is an integer selected from 1 and 2.

In some embodiments, the ratio of the compound of Formula XVIII and theGrignard or lithium reagent is from about 1:1 to about 1:2.0, about 1:1to about 1:1.8, or about 1:1 to about 1.2. Typically, the reaction isconducted in a non-protic organic solvent. In some embodiments, thesolvent is tetrahydrofuran. In some embodiments, the ratio of thecompound of Formula XVIII to the compound of Formula XVII is from about1:1 to about 1:5, about 1:1 to about 1:3, or about 1:1.5 to about 1:2.5.

In some embodiments, a Grignard reagent is used in step (a) and thetemperature is from −30° C. to about room temperature, about −30 toabout 0° C., or about −25 to about −5° C. In some embodiments, a lithiumreagent is used in step (a) and the temperature is from about −80° C. toabout −60° C., or about −78° C.

In some embodiments, the Grignard reagent is isopropyl magnesiumbromide, or adduct thereof.

In some embodiments, R₄ is C₁₋₄ alkyl. In some embodiments, R₄ is C₁₋₃alkyl. In some embodiments, R₄ is methyl or isopropyl. In someembodiments, X₃ is iodo or bromo. In some embodiments, m is 2. In someembodiments, m is 1.

Compounds of Formula XVIII may be known in some cases (see, e.g., Abe,et al., Heterocycles, 2005, 66, 229-240; Korolev, et al., Tet. Lett.2005, 46, 5751-5754; Vasilevsky, Heterocycles, 2003, 60(4), 879-886; andWO 2008/082198, each of which is incorporated herein by reference in itsentirety). In other embodiments, the process further comprises a methodfor preparing a compound of Formula XVIII, comprising protecting acompound of Formula XIX:

wherein:

P₃ is a protecting group;

X₃ is halogen; and

m is an integer selected from 1 and 2.

Di-substituted and mono-substituted compounds of Formula XIX may beknown in some case (see, e.g., WO 2007/043677; Vasilevsky, Heterocycles,2003, 60(4), 879-886; WO 2008/013925; and Huttel, et al., Ann. 1959,625, 55, each of which is incorporated herein by reference in itsentirety). In some embodiments, the process further comprises a methodfor preparing a compound of Formula XIX, comprising reacting a1H-pyrazole with a halogenating agent;

wherein:

X₃ is halogen; and

m is an integer selected from 1 and 2.

In some embodiments, X₃ is iodo or bromo. In some embodiments, thehalogenating agent is selected from N-bromosuccinimide (NBS) orN-iodosuccinimide, wherein X₃ is bromo or iodo.

The intermediates formed from the processes described herein can be usedas appropriate in the other processes described herein.

iii. Preparation of 4-Chloro-7H-pyrrolo[2,3-d]pyrimidine

Compounds of Formula XI are useful intermediates in some of thesynthetic processes described herein. In some embodiments, the presentinvention provides a process for preparing4-chloro-7H-pyrrolo[2,3-d]pyrimidine (XIa), which is a compound ofFormula XI, wherein X₂ is chloro (Scheme 15).

4-Chloro-7H-pyrrolo[2,3-d]pyrimidine (XIa) is synthesized by treating acompound of Formula F-1 with acid. The compound of Formula F-1 can besynthesized by treating a compound of Formula F-2 with a Wittig reagenthaving a glide of formula CH₂OCH₃. The compound of Formula F-2 can beformed starting from commercially available 4,6-dihydroxypyrimidine(compound F-4) by a Vilsmeier Formylation-chlorination to form acompound of Formula F-3, followed by selective ammonolysis to form acompound of Formula F-2.

Accordingly, in some embodiments, the present invention provides aprocess for preparing a compound of Formula XIa:

comprising treating a compound of Formula F-1:

with acid under conditions sufficient to form a compound of Formula D-1.

In some embodiments, the acid is a strong acid. In some embodiments, theacid is aqueous concentrated hydrochloric acid (about 18 M). In someembodiments, the conditions comprising conducting the reacting in asolvent at reflux temperatures. In some embodiments, the reaction iscomplete in about 5 to about 15 hours.

In some embodiments, the process further comprises a process forpreparing a compound of Formula F-1, comprising reacting a compound ofFormula F-2:

with about 1 or more equivalents of a Wittig-type reagent having a ylideof formula —CH₂OCH₃ in the presence of a base.

As used herein, the term “Wittig-type reagent” refers to reagents usedin the Wittig reaction, the Wadsworth-Emmons reaction, and theHorner-Wittig reaction as described in the art (see e.g., Carey andSundberg, Advanced Organic Chemistry, Part B: Reactions and Synthesis,4th ed., Kluwer Academic/Plenum Publishers:New York, pages 111-119(2001); and March, Advanced Organic Chemistry: Reactions, Mechanisms,and Structure, 3rd ed., John Wiley & Sons:New York, pages 845-855(1985), each of which is incorporated herein by references in itsentirety). Exemplative Wittig-type reagents containing a cyanomethyl orcyanomethyl ylide group include, but are not limited to, compounds ofgeneral formula (R′O)₂P(═O)-L-R¹, R″₃P(+)-L(−)-R¹, R″₃P(+)-L-R¹X;R″₂P(═O)-L-R¹, and (R′N)₂P(═O)-L-R¹, wherein R′ is C₁₋₆ alkoxy oroptionally substituted phenyl; R″ is optionally substituted phenyl; L is—CH₂— or —CH—; and R¹ is methoxy; and X is an anion (e.g., halo anion,such as chloride). In some embodiments, the Wittig-type reagent isdiethyl methoxymethyl phosphate. In some embodiments, the reacting ofthe compound of Formula F-1 with the Wittig-type reagent in the presenceof a base. In some embodiments, the base is a strong base. In someembodiments, the base is potassium t-butoxide, sodium t-butoxide, sodiumhydride, sodium ethoxide, sodium hydroxide, potassium carbonate, orsodium carbonate. In some embodiments, the base is an alkali metalalkoxide. In some embodiments, the base is an alkali metal t-butoxide.In some embodiments, the base is potassium t-butoxide. In someembodiments, the olefination of the aldehyde of Formula F-1 with aWittig-type reagent is conducted in an organic solvent, such as THF,under the influence a base, such as potassium tert-butoxide, at atemperature from about 0 to about 5° C. In some embodiments, the base ispresent in about 1 to about 1.2 equivalents, or about 1.05 to about 1.1equivalents, with respect to the compound of Formula F-1. In someembodiments, the Wittig-type reagent is present in about 1 to about 1.2equivalents, or about 1.05 to about 1.1 equivalents with respect to thecompound of Formula F-1. In some embodiments, the Wittig-type reagent is(methoxymethyl)triphenylphosphinium chloride.

In some embodiments, the process further comprises a process forpreparing a compound of Formula F-2, comprising reacting a compound ofFormula F-3:

with about 2 or more equivalents of ammonia in a solvent.

In some embodiments, the solvent is methanol. In some embodiments, theammonia is present in about two equivalents with respect to the compoundof Formula F-2.

In some embodiments, the process further comprises a process forpreparing a compound of Formula F-3, comprising reacting a compound ofFormula F-4:

with a chlorinating agent.

In some embodiments, the chlorinating agent is phosphorous oxychloride.In some embodiments, the chlorinating agent is present in about orgreater than about 2 equivalents, about or greater than about 3equivalents, or about or greater than about 4 equivalents, or from about3 to about 5 equivalents with respect to the compound of Formula F-3.

The intermediates formed from the processes described herein can be usedas appropriate in the other processes described herein.

SPECIFIC EMBODIMENTS

In some embodiments, the present invention provides a process ofpreparing a composition comprising an enantiomeric excess of equal to orgreater than 90% of the (R)-enantiomer of a compound of Formula III′:

comprising:

(a) treating a compound of Formula XI′:

with sodium hydride and N-pivaloyloxymethyl chloride to form a compoundof Formula X′:

(b) treating the compound of Formula X′ with a compound of FormulaXIII′:

in the presence of Pd(triphenylphosphine)₄, potassium carbonate, and asolvent, to form a compound of Formula XII′:

(c) reacting the compound of Formula XII′ under deprotection conditionsto give a compound of Formula IV′:

(d) reacting the compound of Formula IV″ with a compound of FormulaXIV′:

in the presence of 1,8-diazabicyclo[5.4.0]undec-7-ene to give a compoundof Formula II′:

(e) reacting the compound of Formula II′ with hydrogen gas in thepresence of [Rh(COD)₂]CF₃SO₃ and a chiral phosphine ligand selectedfrom:

to form a compound of Formula I′:

and

(f) reacting the compound of Formula I′ under deprotection conditions toform the compound of Formula III′;

wherein * indicates a chiral carbon.

In some embodiments of step (e):

-   -   the solvent is 2,2,2-trifluoroethanol (TFE);    -   the hydrogenation catalyst loading is about 0.005 to about 0.01        mol %;    -   the ratio of the compound of Formula II to the hydrogenation        catalyst is from about 20000/1 to about 10000/1;    -   the hydrogen pressure is from about 7 to about 60 bar;    -   the reacting is run at a temperature from about room temperature        to about 75° C.;    -   the reacting is run until the conversion of the compound of        Formula II to the compound of Formula is about equal to or        greater than 99.5%; and    -   the reacting is from about 10 to about 25 hours.

In some embodiments, the process further comprises preparing thecompound of Formula XI′:

comprising:

(i) reacting a compound of F-4:

with from about three to about five equivalents of POCl₃ in the presenceof about one to about two equivalents of dimethylformamide to form acompound of Formula F-3:

(ii) reacting the compound of F-3 with about two equivalents of ammoniain methanol to form a compound of Formula F-2:

(iii) reacting the compound of Formula F-2 with from about 1 to about1.5 equivalents of a Wittig-type reagent of formula [Ph₃P⁺(CH₂OCH₃)]Cl—,wherein Ph is phenyl, in the presence of from about 1 to about 1.5equivalents of potassium tert-butoxide to form a compound of FormulaF-1:

and

(iv) treating the compound of Formula F-1 with aqueous concentratedhydrochloric acid in tetrahydrofuran at reflux to form compound ofFormula XI′.

In some embodiments, the process further comprises preparing thecompound of Formula XIII′:

comprising:

(i) reacting 1H-pyrazole with N-bromosuccinimide to form a compound ofFormula XIX′;

(ii) protecting the compound of Formula XIX to form a compound ofFormula XVIII′:

and

(iii) reacting the compound of Formula XVIII′ with about one or moreequivalents of a isopropylmagnesium chloride followed by treating withabout one or more equivalents of compound of Formula XVII′:

to form a compound of Formula XIII′.

In some embodiments, the process further comprises preparing thecompound of Formula XIII′:

comprising:

(i) protecting 4-iodo-1H-pyrazole to form a compound of Formula XVIII″:

and

(ii) reacting a compound of Formula XVIII″ with about one or moreequivalents of a isopropylmagnesium chloride in tetrahydrofuran followedby treating with about one or more equivalents of compound of FormulaXVII′:

to form a compound of Formula XIII′.

In further embodiments, the present invention provides a process ofpreparing a composition comprising an enantiomeric excess of the(R)-enantiomer of a compound of Formula III′:

comprising:

(a) treating a compound of Formula XI′:

with sodium hydride and 2-(trimethylsilyl)ethoxymethyl to form acompound of Formula X″:

(b) treating the compound of Formula X″ with a compound of FormulaXIII′:

in the presence of Pd(triphenylphosphine)₄, potassium carbonate, and asolvent, to form a compound of Formula XII″:

(c) reacting the compound of Formula XII″ under deprotection conditionsto form a compound of Formula IV″:

(d) reacting the compound of Formula IV″ with a compound of FormulaD-1′:

under conditions sufficient to form a composition comprising a racemateof a compound of Formula I″:

(e) passing the composition comprising the racemate of the compound ofFormula I″ through a chiral chromatography unit using a mobile phase andcollecting a composition comprising an enantiomeric excess of the(R)-enantiomer of the compound of Formula I″; and

(f) reacting the compound of Formula I″ with lithium tetrafluoroborate,followed by aqueous ammonium hydroxide to form a composition comprisingan enantiomeric excess of the (R)-enantiomer of the compound of FormulaIII′;

wherein * is a chiral carbon.

In other embodiments, the present invention provides a process ofpreparing a composition comprising an enantiomeric excess of the(R)-enantiomer of a compound of Formula III′:

comprising:

(a) treating a composition comprising an enantiomeric excess of the(S)-enantiomer of a compound of Formula I″:

with a compound of Formula D-1′:

in the presence of cesium carbonate in acetonitrile under conditionssufficient to form the racemate of the compound of Formula I″;

(b) passing the composition comprising the racemate of the compound ofFormula I″ through a chiral chromatography unit using a mobile phase andcollecting a composition comprising an enantiomeric excess of the(R)-enantiomer of the compound of Formula I″; and

(c) reacting the compound of Formula I″ with lithium tetrafluoroborate,followed by aqueous ammonium hydroxide to form a composition comprisingan enantiomeric excess of the (R)-enantiomer of the compound of FormulaIII′;

wherein * is a chiral carbon.

In other embodiments, the present invention provides a process ofpreparing a composition comprising an enantiomeric excess of the(R)-enantiomer of a compound of Formula III′:

comprising:

(a) treating a compound of Formula XI′:

with sodium hydride and 2-(trimethylsilyl)ethoxymethyl to form acompound of Formula X″:

(b) treating said compound of Formula X″ with a compound of FormulaXIII′:

in the presence of Pd(triphenylphosphine)₄, potassium carbonate, and asolvent, to form a compound of Formula XII″:

(c) reacting said compound of Formula XII″ under deprotection conditionsto form a compound of Formula IV″:

(d) reacting said compound of Formula IV″ with a compound of FormulaD-1′:

under conditions sufficient to form a composition comprising a racemateof a compound of Formula I″:

(e) passing said composition comprising said racemate of said compoundof Formula I″ through a chiral chromatography unit using a mobile phaseand collecting a composition comprising an enantiomeric excess of the(R)-enantiomer of said compound of Formula I″; and

(f) reacting said compound of Formula I″ with boron trifluoride diethyletherate, followed by aqueous ammonium hydroxide to form a compositioncomprising an enantiomeric excess of the (R)-enantiomer of said compoundof Formula III′;

wherein * is a chiral carbon.

In other embodiments, the present invention provides a process ofpreparing a composition comprising an enantiomeric excess of the(R)-enantiomer of a compound of Formula III′:

comprising:

(a) treating a composition comprising an enantiomeric excess of the(S)-enantiomer of a compound of Formula I″:

with a compound of Formula D-1′:

in the presence of cesium carbonate in acetonitrile under conditionssufficient to form the racemate of the compound of Formula I″;

(b) passing said composition comprising said racemate of said compoundof Formula I″ through a chiral chromatography unit using a mobile phaseand collecting a composition comprising an enantiomeric excess of the(R)-enantiomer of said compound of Formula I″; and

(c) reacting said compound of Formula I″ with boron trifluoride diethyletherate, followed by aqueous ammonium hydroxide to form a compositioncomprising an enantiomeric excess of the (R)-enantiomer of said compoundof Formula III′;

wherein * is a chiral carbon.

In other embodiments, the present invention provides a process ofpreparing a composition comprising an enantiomeric excess of the(R)-enantiomer of a compound of Formula III′:

comprising: reacting said compound of Formula I″:

with boron trifluoride diethyl etherate, followed by aqueous ammoniumhydroxide to form a composition comprising an enantiomeric excess of the(R)-enantiomer of said compound of Formula III′; wherein * is a chiralcarbon.

In other embodiments, the present invention provides a process forpreparing(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt comprising reacting(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilewith phosphoric acid in the presence of 2-propanol and dichloromethane.

In other embodiments, the present invention provides a method ofpurifying(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt comprising recrystallizing(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt from a solvent mixture comprising methanol, 2-propanol,and n-heptane. In some embodiments, the 2-propanol and n-heptane areadded to a mixture of(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt in methanol.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of noncriticalparameters which can be changed or modified to yield essentially thesame results.

EXAMPLES

[4-(1H-Pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate (5)

To a oven dried 3 L 4-neck round bottom flask equipped with a stirringbar, a septa, a thermocouple, a 500 mL addition funnel and the nitrogeninlet was charged sodium hydride (NaH, 60 wt % in mineral oil, 32.82 g,0.82 mol, 1.20 equiv) and anhydrous 1,2-dimethoxyethane (DME, 500 mL,4.8 mol) and the resulting mixture was cooled to 0-3° C. To a oven dried1 L round bottom flask was charged 4-chloro-7H-pyrrolo[2,3-d]pyrimidine(1, 105.0 g, 0.684 mol) and 1,2-dimethoxyethane (DME, 750 mL, 7.2 mol)and the resulting slurry was then portion wise added to the suspensionof sodium hydride in DME via large bore canula over 30 min at 5-12° C.The resulting reaction mixture was heterogeneous. Following theaddition, the cold bath was removed and the mixture was gradually warmedto room temperature and allowed to stir at room temperature for 1 hourbefore being cooled to 0-5° C. Chloromethyl pivalate (pivaloyloxymethylchloride, POM-Cl, 112 ml, 0.752 mol, 1.1 equiv) was added dropwise intothe reaction mixture over 30 minutes with stirring at 0-5° C. Theaddition of chloromethyl pivalate was mildly exothermic and the reactiontemperature went up to as high as 14° C. After addition of chloromethylpivalate, the cooling bath was removed and the reaction mixture wasallowed to return to room temperature and stirred at room temperaturefor 90 min. When the reaction was deemed complete as confirmed by TLCand LCMS, the reaction was carefully quenched with water (100 mL). Andthis quenched reaction mixture, which contains crude POM-protectedchlorodeazapurine (2), was directly used in the subsequent Suzukicoupling reaction without further work-up and purification.

To the quenched reaction mixture, which contains crude POM-protectedchlorodeazapurine (2) made as described above,4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (3, 200 g,0.75 mol, 1.10 equiv) and potassium carbonate (K₂CO₃, 189 g, 1.37 mol,2.0 equiv) were added at room temperature. The resulting mixture wasdegassed by passing a stream of nitrogen through the solution for 15minutes before being treated withtetrakis(triphenylphosphine)palladium(0) (Pd(PPh₃)₄, 7.9 g, 0.68 mmol,0.01 equiv) and the resulting reaction mixture was heated at reflux(about 82° C.) for 10 h. When the reaction was deemed complete asconfirmed by TLC (1:1 hexanes/ethyl acetate) and LCMS, the reactionmixture was cooled down to room temperature and diluted with ethylacetate (2 L) and water (1 L). The two layers were separated, and theaqueous layer was extracted with ethyl acetate (EtOAc, 500 mL). Thecombined organic layers were washed with water (2×1 L) and brine (1 L)before being concentrated under reduced pressure to afford crude{4-[1-(1-ethoxyethyl)-1H-pyrazol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methylpivalate (4) as a pale-yellow oil, which was directly used in thesubsequent de-protection reaction without further purification.

To a solution of crude{4-[1-(1-ethoxyethyl)-1H-pyrazol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methylpivalate (4) in THF (1 L, 12.3 mol), a 4 N aqueous HCl solution (500 mL)was added at room temperature. The resulting reaction mixture wassubsequently stirred at room temperature for 5 h. When the reaction wasdeemed complete as confirmed by TLC and LCMS, the reaction mixture wascooled to 0-5° C. before pH was adjusted to 9-10 with a 1 M aqueoussodium hydroxide (NaOH) solution (2 L). The mixture was concentratedunder reduced pressure to remove most of THF and the resultingsuspension was stirred at room temperature for 2 h. The solids werecollected by filtration, washed with water (3×500 mL), and dried invacuum to afford crude[4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate (5,157.5 g, 204.43 g theoretical, 77% yield for three steps) as white tooff-white solids, which was found to be sufficiently pure (>98 area % byHPLC) to do the subsequent reaction without further purification. For 5:¹H NMR (DMSO-d₆, 400 MHz) δ ppm 13.42 (br s, 1H), 8.76 (s, 1H), 8.67 (s,1H), 8.33 (s, 1H), 7.68 (d, 1H, J=3.8 Hz), 7.11 (d, 1H, J=3.8 Hz), 6.21(s, 2H), 1.06 (s, 9H); ¹³C NMR (DMSO-d₆, 100 MHz) δ ppm 177.74, 152.31,152.09, 151.91, 139.52, 130.39, 120.51, 113.93, 101.91, 67.26, 38.98,27.26; C₁₅H₁₇N₅O₂ (MW, 299.33), LCMS (EI) m/e 300 (M⁺+H).

Cyanatobenzene (6)

To a oven dried 500 mL 3-neck round bottom flask equipped with aoverhead stirring, a septa, a thermocouple and the nitrogen inlet,phenol (20.0 g, 0.210 mol), diethyl ether (Et₂O, 290 mL) and cyanicbromide (BrCN, 23.0 g, 0.210 mol, 1.0 equiv) were added at roomtemperature. The resulting solution was cooled down to 0-3° C. beforetriethylamine (TEA, 61.9 mL, 0.442 mol, 2.1 equiv) was added dropwisevia syringe over 25 min. The addition of triethylamine to reactionmixture was mildly exothermic and the reaction temperature went up to ashigh as 15° C. After addition of triethylamine, the reaction mixturebecame a white slurry which was stirred vigorously at 0° C. for 2 h at5-15° C. When the reaction was deemed complete as confirmed by TLC andLCMS, the reaction mixture was diluted with pentane (150 mL, 1.30 mol).The precipitated triethylamine hydrochloride was filtered off and thesalt was washed with diethyl ether and pentane (1 to 1 by volume, 200mL). The filtrate was then concentrated under reduced pressure to removethe majority of the solvent, and the residue, which contains the crudecyanatobenzene (6), was directly used in the subsequent reaction withoutfurther purification assuming the theoretical yield.

3-Cyclopentylpropiolonitrile (8)

To a oven dried 500 mL 3-neck round bottom flask equipped with a stirbar, a nitrogen inlet, a 125 mL addition funnel and a thermocouple,cyclopentylacetylene (7, 15.0 g, 0.143 mol) and anhydroustetrahydrofuran (THF, 170 mL, 2.10 mol) were added at room temperature.The resulting solution was then cooled to −78° C. before a solution of2.5 M n-butyllithium in hexane (63.1 mL, 0.158 mol, 1.1 equiv) was addeddropwise over 25 min. The resulting lithium cyclopentylacetylenesolution was stirred at −78° C. for 15 minutes before a solution ofcrude cyanatobenzene (6, 25.0 g, 0.210 mol, 1.5 equiv) in anhydroustetrahydrofuran (THF, 30.0 mL, 0.400 mol) was added dropwise via acanula at −78° C. The resulting reaction mixture was stirred at −78° C.for an additional 10 min before the cooling batch was removed and thereaction mixture was allowed to gradually warm to room temperature andstirred at room temperature for 1-2 h. When the reaction was deemedcomplete, the reaction mixture was quenched with a 6 N aqueous sodiumhydroxide solution (NaOH, 200 mL) and a 20% aqueous brine solution (200mL). The aqueous solution was treated with ethyl acetate (EtOAc, 200 mL)before the two layers were separated. The organic layer was dried overmagnesium sulfate (MgSO₄), filtered, and concentrated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,0 to 5% ethyl acetate/hexane gradient elution) to afford3-cyclopentylpropiolonitrile (8, 14.3 g, 17.0 g theoretical, 84% yieldfor two steps) as a yellow to orange oil. For 8: ¹H NMR (DMSO-d₆, 400MHz) δ 2.97 (m, 1H), 1.97 (m, 2H), 1.64 (m, 4H), 1.56 (m, 2H).

4-(1-(2-Cyano-1-cyclopentylvinyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (9)

To a 500 mL round bottom flask equipped with a stir bar and the nitrogeninlet was charged 3-cyclopentylpropiolonitrile (8, 8.50 g, 0.0713 mol,1.52 equiv), N,N-dimethylformamide (DMF, 84 mL, 1.08 mol) and[4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate (5,14.0 g, 0.0468 mol) and solid potassium carbonate (K₂CO₃, 0.329 g,0.00238 mol, 0.05 equiv) at room temperature. The resulting reactionmixture was then stirred at room temperature for 60 min. When TLC andHPLC showed that the reaction was deemed complete, the reaction mixturewas quenched with 20% aqueous brine (75 mL) and the resulting solutionwas extracted with ethyl acetate (EtOAc, 3×75 mL). The combined organicextracts were washed with 20% aqueous brine (75 mL), dried overmagnesium sulfate (MgSO₄), filtered, and concentrated under reducedpressure. The residue was purified by flash chromatography (SiO₂, 0 to20% ethyl acetate/hexane gradient elution) to afford4-(1-(2-cyano-1-cyclopentylvinyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (9, 16.4 g, 19.6 g theoretical, 83.7% yield) as white solids.For 9: ¹H NMR (DMSO-d₆, 300 MHz) δ 9.09 (s, 1H), 8.84 (s, 1H), 8.63 (s,1H), 7.78 (d, 1H, J=3.8 Hz), 7.17 (d, 1H, J=3.8 Hz), 6.24 (s, 2H), 5.82(s, 1H), 3.55 (m, 1H), 1.92 (m, 2H), 1.59 (br m, 6H), 1.06 (s, 9H);C₂₃H₂₆N₆O₂ (MW, 418.49), LCMS (EI) m/e 419 (M⁺+H).

(Z)-(4-(1-(3-Amino-1-cyclopentyl-3-oxoprop-1-enyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (10)

To a 200 ml round bottom flask equipped with a stir bar and the nitrogeninlet was charged4-(1-(2-cyano-1-cyclopentylvinyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (9, 8.00 g, 0.0191 mol), trifluoroacetic acid (TFA, 40.6 mL,0.528 mol) and concentrated sulfuric acid (H₂SO₄, 3.77 mL, 0.0707 mol)at room temperature. The resulting reaction mixture was stirred at roomtemperature for 60 min. When TLC and HPLC showed that the reaction wasdeemed complete, the reaction mixture was quenched with water (30.1 mL,1.67 mol). The quenched reaction mixture was stirred at room temperaturefor 30 min before being cooled to 0-5° C. The cold solution was thentreated with a 3 N sodium hydroxide aqueous solution (NaOH, 223 mL) toadjust pH to 8 before being treated with ethyl acetate (EtOAc, 200 mL).The two layers were separated, and the aqueous was then extracted withethyl acetate (EtOAc, 2×50 mL). The combine organic extracts were washedwith a saturated aqueous NaCl (100 mL), dried over magnesium sulfate(MgSO₄), filtered, and concentrate under reduced pressure. The residuewas purified by flash chromatography (SiO₂, 0 to 100% ethylacetate/hexane gradient elution) to provide(Z)-(4-(1-(3-amino-1-cyclopentyl-3-oxoprop-1-enyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (10, 6.79 g, 8.34 g theoretical, 81.4% yield) as light yellowsolids. For 10: ¹H NMR (DMSO-d₆, 300 MHz) δ 8.77 (s, 1H), 8.68 (s, 1H),8.41 (s, 1H), 7.71 (d, 1H, J=3.8 Hz), 7.51 (br. s, 1H), 7.09 (br. s,1H), 7.05 (d, 1H, J=3.8 Hz), 6.22 (s, 2H), 5.97 (s, 1H), 3.27 (m, 1H),1.77 (m, 2H), 1.54 (m, 6H), 1.06 (s, 9H); C₂₃H₂₈N₆O₃ (MW, 436.51), LCMS(EI) m/e 437 (M⁺+H).

(4-(1-(3-Amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (11)

To a 25 ml round bottom flask equipped with a stir bar was charged(Z)-(4-(1-(3-amino-1-cyclopentyl-3-oxoprop-1-enyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (10, 1.15 g, 2.63 mmol), tetrahydrofuran (THF, 20.0 ml, 246mmol) and 10% palladium on carbon (50% wet, 130 mg) at room temperature.The resulting reaction mixture was degassed three times back fillingwith hydrogen gas each time before the hydrogenation reaction wasconducted under a steady stream of hydrogen gas released by a hydrogenballoon. The reaction was checked after 17 h and was found to becomplete. The reaction mixture was then filtered through a Celite bed toremove the catalyst and the Celite bed was rinsed with a small volume oftetrahydrofuran (THF). The combined filtrates were concentrated underreduced pressure to afford the crude(4-(1-(3-amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (11, 1.15 g, 1.153 g theoretical, 99% yield) as a yellow tobrown oil, which solidified upon standing in vacuum at room temperature.This crude product (11) was found to be pure enough (>98% by HPLC) to dothe following reaction without further purification. For 11: ¹H NMR(DMSO-d₆, 300 MHz) δ ppm 8.73 (s, 1H), 8.60 (s, 1H), 8.27 (s, 1H), 7.70(d, 1H, J=3.8 Hz), 7.32 (bs, 1H), 7.09 (d, 1H, J=3.8 Hz), 6.75 (bs, 1H),6.21 (s, 2H), 4.56 (td, 1H, J=4.0, 9.8 Hz), 2.86 (dd, 1H, J=10.5, 5.6Hz), 2.63 (dd, 1H, J=4.0, 15.3 Hz), 2.32 (m, 1H), 1.77 (m, 1H),1.56-1.19 (m, 7H), 1.06 (s, 9H); LCMS (EI) m/e 439 (M⁺+H); C₂₃H₃₀N₆O₃(MW, 438.52), LCMS (EI) m/e 439 (M⁺+H).

(R)-(4-(1-(3-Amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-11) and(S)-(4-(1-(3-Amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((S)-11)

General screening procedure for asymmetric hydrogenation using thesubstrate,(Z)-(4-(1-(3-amino-1-cyclopentyl-3-oxoprop-1-enyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (10) to afford optically enriched product,(4-(1-(3-amino-1-cyclopentyl-3-oxopropyl]-1H-pyrazol-4-yl}-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-11 or (S)-11): A 300 mL-volume autoclave with glass vial(20 mL) was charged with the substrate (10), the catalyst (metal,ligand, and catalyst precursor), and oxygen-free solvent (4-6 mL) undernitrogen. This autoclave was charged with hydrogen gas to the desiredpressure and stirred at room temperature or heated with oil bath. Afterhydrogen gas was released, the reaction mixture was concentrated underreduced pressure. The residue was purified by eluting through a silicagel pad using a mixture of ethyl acetate and methanol (v/v=9/1) toafford product,(4-(1-(3-amino-1-cyclopentyl-3-oxopropyl]-1H-pyrazol-4-yl}-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-11 or (S)-11), for chemical conversion (HPLC and chiralHPLC), LC/MS and NMR spectroscopy and enantiomeric excess (% ee bychiral HPLC) determination.

The determination of enantiomeric excess (% ee) of the product wascarried out by chiral HPLC analysis. A Chiralpak® IA column was used.The mobile phase was a mixture of hexane and ethanol (v/v=90/10). Theflow rate was 1 mL/min and UV detection wavelength was set at 254 nm.The substrate (10), undesired enantiomer ((S)-11, 1^(st) peak) anddesired enantiomer ((R)-11, 2^(nd) peak) were well-resolved at retentiontimes 46 min, 36 min, and 38 min respectively.

For (R)-11 or (S)-11: ¹H NMR (DMSO-d₆, 300 MHz) δ ppm 8.73 (s, 1H), 8.60(s, 1H), 8.27 (s, 1H), 7.70 (d, 1H, J=3.8 Hz), 7.32 (bs, 1H), 7.09 (d,1H, J=3.8 Hz), 6.75 (bs, 1H), 6.21 (s, 2H), 4.56 (td, 1H, J=4.0, 9.8Hz), 2.86 (dd, 1H, J=10.5, 5.6 Hz), 2.63 (dd, 1H, J=4.0, 15.3 Hz), 2.32(m, 1H), 1.77 (m, 1H), 1.56-1.19 (m, 7H), 1.06 (s, 9H); LCMS (EI) m/e439 (M⁺+H); C₂₃H₃₀N₆O₃ (MW, 438.52), LCMS (EI) m/e 439 (M⁺+H).

The following table summarizes analyses and reaction conditions for thisasymmetric hydrogenation.

Major Metal/Ligand/ H₂ Conversion Enantiomer Catalyst Temp. PressureTime (HPLC % (R)- or (S)- Precursor Solvent (° C.) (Bar) (h) Area %) ee11 Rh(COD)(SSRR- CF₃CH₂OH 23 40 20 99 66 (S)-11 TangPhos) (1^(st) peak)(BF₄) Rh(COD)(SSRR- CH₂Cl₂ 23 40 20 91 92 (S)-11 TangPhos) (1^(st) peak)(BF₄) Rh(COD)(SSRR- CF₃CH₂OH 23 10 20 99 91 (S)-11 TangPhos) & CH₂Cl₂(1^(st) peak) (BF₄) Rh(COD)(+)- DuanPhos) MeOH 23 40 20 94 66 (S)-11(BF₄) (1^(st) peak) Rh(COD)(+)- DuanPhos) CF₃CH₂OH 23 40 20 99 61 (S)-11(BF₄) (1^(st) peak) Ru(R-C3- TunePhos) MeOH 50 50 2 84 87 (R)-11(CF₃CO₂)₂ (2^(nd) peak) Ru(R-C3- TunePhos) CF₃CH₂OH 50 50 2 99 88 (R)-11(CF₃CO₂)₂ & MeOH (2^(nd) peak) Ru(COD)(SL-A153-1) MeOH 30 50 21 100 98(R)-11 (CF₃CO₂)₂ (2^(nd) peak) Rh(COD)2 (SL-W008-1) MeOH 30 50 21 100 94(R)-11 (CF₃SO₃) (2^(nd) peak)

Structures of chiral phosphine ligands used in this study are listedbelow.

Representative preparative asymmetric hydrogenation procedure andproduct chiral purity upgrade by crystallization are described below.

(S)-(4-(1-(3-Amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((S)-11)

A solution of(4-{1-[(1Z)-3-amino-1-cyclopentyl-3-oxoprop-1-en-1-yl]-1H-pyrazol-4-yl}-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (10, 215 mg) in a mixture of methylene chloride (CH₂Cl₂, 12.5mL) and trifluoroethanol (CF₃CH₂OH, 0.25 mL) in a pressure glass tubewas treated with the catalyst Rh(COD)(SSRR-TangPhos)BF₄ (8.8 mg) undernitrogen before the reaction mixture was pressurized with hydrogen gasto 40 bar pressure. The reaction mixture was stirred at 50° C. underthis hydrogen pressure for 20 h. When HPLC analysis showed that thesubstrate was completely consumed, the reaction mixture was cooled downto room temperature. The enantiomeric excess of the reaction mixture wasdetermined to be 88% ee (94% of the first peak, (S)-11; 6% of the secondpeak, (R)-11) by chiral HPLC analysis. The reaction mixture was filteredthrough a thin silica gel pad and the pad was washed with methylenechloride (5 mL). The filtrate was then concentrated under reducedpressure to dryness. The resultant foamy solid (180 mg) was charged witha mixture of heptane (5 mL) and ethyl acetate (EtOAc, 5 mL). Whitesolids precipitated out upon stirring at 20° C. The slurry was stirredat 20° C. for 16 h. The solid was collected by filtration and the chiralHPLC analysis for the collected solids (52 mg) showed a 66.0% ofenantiomeric excess favoring the first peak (83.0% of the first peak,(S)-11; 17.0% of the second peak, (R)-11). The filtrate obtained wasthen evaporated to dryness. The resultant oil (108 mg) was analyzed bychiral HPLC and showed a 99.6% of enantiomeric excess favoring the firstpeak (99.83% of the first peak, (S)-11; 0.17% of the second peak,(R)-11). This result showed in principle that the optical purity of theasymmetric hydrogenation product can be significantly enhanced byselective removal of the minor enantiomer by precipitation of the solidusing a suitable solvent system such as ethyl acetate/heptane asdescribed.

(R)-(4-(1-(3-Amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-11)

A solution of(4-{1-[(1Z)-3-amino-1-cyclopentyl-3-oxoprop-1-en-1-yl]-1H-pyrazol-4-yl}-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate, (10, 500 mg) in methanol (MeOH, 8.0 mL) in a pressure glasstube was treated with the catalyst Ru(COD)(SL-A153-1)(CF₃CO₂)₂ (6.6 mg)under nitrogen before the reaction mixture was pressurized with hydrogengas to 50 bar pressure. The reaction mixture was stirred at 30° C. underthis hydrogen pressure for 21 h. When HPLC analysis showed that thesubstrate was completely consumed, the reaction mixture was cooled downto room temperature. The enantiomeric excess of the reaction mixture wasdetermined to be 98% ee (99% of the second peak, (R)-11; 1% of the firstpeak, (S)-11) by chiral HPLC analysis. The reaction mixture was thenfiltered through a thin silica gel pad and the pad was washed withmethanol (5 mL). The filtrate was then concentrated under reducedpressure to dryness. The resultant foamy solid (470 mg) was analyzed bychiral HPLC analysis and result showed a 98.0% of enantiomeric excessfavoring the second peak (99.0% of the second peak, (R)-11; 1.0% of thefirst peak, (S)-11).

(R)-(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-12)

Method A. To a 50 mL round bottom flask equipped with a stir bar and thenitrogen inlet was charged(R)-(4-(1-(3-amino-1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-11, 413 mg, 0.942 mmol), N,N-dimethylformamide (DMF, 10mL, 129 mmol) and triethylamine (TEA, 0.525 mL, 3.77 mmol, 4.0 equiv) atroom temperature. The resulting mixture was then cooled to 0-5° C. in anice bath before trichloroacetyl chloride (0.315 mL, 2.82 mmol, 3.0equiv) was added drop wise via a syringe at room temperature. Theresulting reaction mixture was stirred at 0-5° C. for 90 min. When TLCand HPLC showed that the reaction was deemed complete, the reactionmixture was treated with ethyl acetate (EtOAc, 25 mL) and 20% aqueousbrine (20 mL). The two layers were separated, and the aqueous layer wasextracted with ethyl acetate (EtOAc, 2×25 mL). The combined organicextracts were washed with 20% aqueous brine (35 mL), dried overmagnesium sulfate (MgSO₄), filtered, and concentrated under reducedpressure. The residual brown oily crude product was purified by flashchromatography (SiO₂, 0 to 50% ethyl acetate/hexane gradient elution) toafford(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-12, 278 mg, 396.1 mg theoretical, 70.2% yield) as thelight oil, which was solidified upon standing at room temperature invacuum. For (R)-12: achiral purity (99.1 area % by HPLC detected at 220nm); chiral purity (99.6 area % by chiral HPLC; 99.2% ee); ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.84 (s, 1H), 8.78 (s, 1H), 8.39 (s, 1H), 7.74(d, 1H, J=3.7 Hz,), 7.11 (d, 1H, J=3.8 Hz), 6.23 (s, 2H), 4.53 (ddd, 1H,J=9.9, 9.6, 4.2 Hz), 3.26 (dd, 1H, J=17.4, 9.9 Hz), 3.19 (dd, 1H,J=17.2, 4.3 Hz), 2.41 (m, 1H), 1.87-1.13 (m, 8H), 1.07 (s, 9H);C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e 421.4 (M⁺+H).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base)

Method A. To a 25 ml round bottom flask equipped with a stir bar and thenitrogen inlet was charged(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-12, 278 mg, 0.661 mmol) and methanol (MeOH, 2.50 mL, 37.0mmol) at room temperature. The resulting homogeneous reaction solutionwas then treated with a 0.10 M aqueous sodium hydroxide solution (NaOH,1.5 mL, 0.15 mmol, 2.3 equiv) at room temperature. The resultingreaction mixture was stirred at room temperature for 22 hours. When thereaction was deemed complete, the reaction mixture was diluted with 20%aqueous brine (10 mL) and ethyl acetate (EtOAc, 25 mL). The two layerswere separated, and the aqueous layer was extracted with ethyl acetate(EtOAc, 25 mL). The combined organic fractions were dried over magnesiumsulfate (MgSO₄), filtered, and concentrated under reduced pressure. Theresidue was purified by flash chromatography (SiO₂, 0 to 100% ethylacetate/hexane gradient elution) to afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base, 188 mg, 202.5 mg theoretical, 92.8% yield) as acolorless oil, which solidified upon standing at room temperature invacuum. For (R)-13 (free base): ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 12.1(bs, 1H), 8.80 (d, 1H, J=0.42 Hz), 8.67 (s, 1H), 8.37 (s, 1H), 7.59 (dd,1H, J=2.34, 3.51 Hz), 6.98 (dd, 1H, J=1.40, 3.44 Hz), 4.53 (td, 1H,J=19.5, 4.63 Hz), 3.26 (dd, 1H, J=9.77, 17.2 Hz), 3.18 (dd, 1H, J=4.32,17.3 Hz), 2.40 (m, 1H), 1.79 (m, 1H), 1.65 to 1.13 (m, 7H); C₁₇H₁₈N₆(MW,306.37) LCMS (EI) m/e 307 (M⁺+H).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt ((R)-14, phosphate)

To a solution of(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base, 572 g, 1.87 mol) in isopropanol (IPA, 8 L) at 60-65°C. was added a solution of phosphoric acid (186.2 g, 1.9 mol, 1.10equiv) in isopropanol (1.6 L). No exotherm was observed while adding asolution of phosphoric acid, and a precipitate was formed almostimmediately. The resulting mixture was then heated at 76° C. for 1.5hours, then cooled gradually to ambient temperature and stirred at roomtemperature for overnight. The mixture was filtered and the solids werewashed with a mixture of heptane and isopropanol (1/1, v/v, 3 L) beforebeing transferred back to the original flask and stirred in heptane (8L) for one hour. The solids were collected by filtration, washed withheptane (1 L), and dried in a convection oven in vacuum at 40° C. to aconstant weight to afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt ((R)-14, phosphate, 634.2 g, 755 g theoretical, 84%yield) as white to off-white crystalline solids. For (R)-14 (phosphate):mp. 197.6° C.; ¹H NMR (DMSO-d₆, 500 MHz) δ ppm 12.10 (s, 1H), 8.78 (s,1H), 8.68 (s, 1H), 8.36 (s 1H), 7.58 (dd, 1H, J=1.9, 3.5 Hz), 6.97 (d,1H, J=3.6 Hz), 4.52 (td, 1H, J=3.9, 9.7 Hz), 3.25 (dd, 1H, J=9.8, 17.2Hz), 3.16 (dd, 1H, J=4.0, 17.0 Hz), 2.41, (m, 1H), 1.79 (m, 1H), 1.59(m, 1H), 1.51 (m, 2H), 1.42 (m, 1H), 1.29 (m, 2H), 1.18 (m, 1H); ¹³C NMR(DMSO-d₆, 125 MHz) δ ppm 152.1, 150.8, 149.8, 139.2, 131.0, 126.8,120.4, 118.1, 112.8, 99.8, 62.5, 44.3, 29.1, 29.0, 24.9, 24.3, 22.5;C₁₇H₁₈N₆(MW, 306.37 for free base) LCMS (EI) m/e 307 (M⁺+H, base peak),329.1 (M⁺+Na).

4-(1H-Pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(17)

To a suspension of sodium hydride (NaH, 60 wt % oil disposition, 4.05 g,101.3 mmol, 1.54 equiv) in 1,2-dimethoxyethane (DME, 20.0 mL, 192.4mmol) at 0-5° C. (ice bath) was added 4-chloropyrrolo[2,3-d]pyrimidine(1, 10.08 g, 65.6 mmol) in 1,2-dimethoxyethane (DME, 80.0 mL, 769.6mmol) slowly so that the temperature was kept at below 5° C. (−7° C. to5° C.). A large amount of gas was evolved immediately after the solutionof substrate (1) was introduced. The resulting reaction mixture wasstirred at 0-5° C. for 30 min before trimethylsilylethoxymethyl chloride(SEM-Cl, 12.56 g, 75.3 mmol, 1.15 equiv) was added slowly while thereaction temperature was maintained at below 5° C. After the addition,the reaction was stirred at 0° C. for 1 h before being warmed to roomtemperature for 23 h. When the HPLC and TLC showed that the reaction wasdeemed complete, the reaction mixture was quenched with water (46 mL) atroom temperature, and the quenched reaction mixture, which contains thedesired product (15), was carried into the next Suzuki coupling reactiondirectly without further work-up and purification.

To the quenched reaction mixture, which contains crude4-chloro-7-[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidine(15, 18.63 g, 65.64 mmol) from previous reaction as described above, wasadded 1,2-dimethoxyethane (DME, 38 mL), powder potassium carbonate(K₂CO₃, 23.56 g, 170.5 mmol, 2.6 equiv),1-(1-ethoxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(3, 18.60 g, 69.89 mmol, 1.06 equiv) at room temperature. The resultingmixture was degassed four times backfilling with nitrogen gas each timebefore being treated with tetrakis(triphenylphosphine)palladium(0)(Pd(PPh₃)₄, 244.2 mg, 0.21 mmol, 0.003 equiv) at room temperature. Theresulting reaction mixture was degassed four times backfilling withnitrogen gas each time before being warmed to 80° C. for 4-8 h. When TLCand HPLC showed that the reaction was deemed complete, the reactionmixture was gradually cooled to room temperature and filtered through ashort bed of Celite (10 g). The Celite bed was washed with ethyl acetate(EtOAc, 20 mL). The two layers of the filtrate were separated, and theaqueous layer was extracted with ethyl acetate (EtOAc, 2×30 mL). Thecombined organic extracts were washed with saturated aqueous NaClsolution (20 mL), dried over magnesium sulfate (MgSO₄), and concentratedunder reduced pressure. The residue, which contains the crude desiredSuzuki coupling product (16), was then transferred to a 500 mL roundbottom flask with THF (22 mL) for subsequent de-protection reactionwithout further purification.

A solution of crude Suzuki coupling product (16) in THF (22 mL) wastreated with water (108 mL) and a solution of 10% aqueous HCl preparedby mixing 19.6 mL of concentrated HCl with 64 mL of H₂O at roomtemperature. The resulting reaction mixture was stirred at roomtemperature for 4-6 h. When TLC and HPLC showed the de-protectionreaction was deemed complete, a 30% aqueous sodium hydroxide (NaOH)solution prepared by dissolving 10.4 g of NaOH in 21.0 mL of H₂O wasadded slowly to the reaction mixture while maintaining the temperaturebelow 25° C. The solid gradually dissolved and re-precipitated after 10min. The mixture was stirred at room temperature for 1-2 h before thesolids were collected by filtration and washed with H₂O (50 mL). The wetcake was transferred to a 250 mL three-necked flask and treated withacetonitrile (MeCN, 112 mL) at room temperature. The mixture was heatedto reflux for 2 h before being cooled gradually to room temperature andstirred at room temperature for 1 h. The solids were collected byfiltration, washed with MeCN (36 mL) and dried at 40-45° C. in a vacuumoven to afford4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(17, 15.3 g, 20.7 g theoretical, 73.9% yield for three steps) as whitecrystalline solids (99.4 area % by HPLC). For 17: ¹H NMR (DMSO-d₆, 400MHz) δ ppm 13.41 (bs, 1H), 8.74 (s, 1H), 8.67 (bs, 1H), 8.35 (bs, 1H),7.72 (d, 1H, J=3.7 Hz), 7.10 (d, 1H, J=3.7 Hz), 5.61 (s, 2H), 3.51 (t,2H, J=8.2 Hz), 0.81 (t, 2H, J=8.2 Hz), 0.13 (s, 9H); C₁₅H₂₁N₅OSi (MW,315.45), LCMS (EI) m/e 316 (M⁺+H).

Methyl 3-cyclopentylpropiolate (18)

To a stirred solution of cyclopentylacetylene (7, 17.49 mL, 150.0 mmol)in anhydrous tetrahydrofuran (THF, 200 mL, 2466 mmol) at −78° C. wasadded 2.50 M of n-butyllithium in hexane (66.0 mL, 165 mmol, 1.1 equiv).The resulting milky suspension was stirred at −78° C. for 30 min. Methylchloroformate (17.6 mL, 225 mmol, 1.5 equiv) was then added. Thereaction mixture became a clear solution. The cooling bath was thenremoved, and the reaction mixture was allowed to warm to roomtemperature and stirred at room temperature for 1 h. The reactionmixture became a suspension again. When TLC (5% EtOAc/hexane, KMnO4stain) showed the reaction was deemed complete, the reaction mixture wasquenched with saturated aqueous NH₄Cl solution (150 mL) and extractedwith diethyl ether (Et₂O, 2×200 mL). The combined organic layers werewashed with saturated aqueous NaCl solution, dried over magnesiumsulfate (MgSO₄), filtered and concentrated under the reduced pressure.The residue was distilled under vacuum (99-101° C./16 mbar) to affordmethyl 3-cyclopentylpropiolate (18, 21.856 g, 22.83 g theoretical, 96%yield) as a colorless oil. For 18: ¹H NMR (CDCl₃, 400 MHz) δ ppm 3.74(s, 3H), 2.73 (m, 1H), 1.95 (m, 2H), 1.72 (m, 4H), 1.57 (m, 2H).

(Z)-3-Cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylonitrile(19)

To a stirred solution of4-(1H-pyrazol-4-yl)-7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidine(17, 7.260 g, 23.01 mmol) and 3-cyclopentylprop-2-ynenitrile (8, 6.140g, 34.52 mmol, 1.5 equiv) in N,N-Dimethylformamide (DMF, 40.0 mL, 516mmol) at room temperature was added solid potassium carbonate (K₂CO₃,318 mg, 2.30 mmol, 0.1 equiv). The resulting reaction mixture wasstirred at room temperature for 30 min. When LCMS showed the reactionwas deemed complete, the reaction mixture was quenched with water (80mL), extracted with EtOAc (2×150 mL). The combined organic layers werewashed with water (80 mL) and brine (50 mL), dried over magnesiumsulfate (MgSO₄), filtered and concentrated under reduced pressure. Theresidue was purified by flash chromatography (SiO₂, 0-30% EtOAc/hexanegradient elution) to give(Z)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylonitrile(19, 8.256 g, 10.0 g theoretical, 82.6% yield) as a colorless syrup. For19: ¹H NMR (CDCl₃, 300 MHz) δ ppm 9.15 (bs, 1H), 8.96 (s, 1H), 8.56 (s,1H), 7.51 (d, 1H, J=3.5 Hz), 6.93 (d, 1H, J=3.5 Hz), 5.75 (s, 2H), 5.29(s, 1H), 3.62 (m, 1H), 3.60 (t, 2H, J=8.2 Hz), 2.16 (m, 2H), 1.81 (m,4H), 1.59 (m, 2H), 0.98 (t, 2H, J=8.2 Hz), 0.00 (s, 9H); C₂₃H₃₀N₆OSi(MW, 434.61), LCMS (EI) m/e 435.2 (M⁺+H).

(R)-3-Cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-20) and(S)-3-Cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((S)-20)

General screening procedure for asymmetric hydrogenation using thesubstrate,(Z)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylonitrile(19), to afford optically enriched product,3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-20 or (S)-20): A 300 mL-volume autoclave with glass vial (20 mL)was charged with the substrate (19), the catalyst (metal, ligand, andcatalyst precursor), and oxygen-free solvent (4-6 mL) under nitrogen.This autoclave was charged with hydrogen gas to the desired pressure andstirred at room temperature or heated with oil bath. After hydrogen gaswas released, the reaction mixture was concentrated under reducedpressure. The residue was purified by eluting through a silica gel padusing a mixture of ethyl acetate and methanol (v/v=9/1) to affordproduct,3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-20 or (S)-20), for chemical conversion (HPLC and chiral HPLC),LC/MS and NMR spectroscopy and enantiomeric excess (% ee by chiral HPLC)determination.

The determination of enantiomeric excess (% ee) of the product wascarried by chiral HPLC analysis. A chiral HPLC method was developedusing a ChiralcelChiralcel® OD-H column (4.6×250 mm, 5 μm), purchasedfrom Chiral Technologies, Inc., packed with a silicagel coated withcellulose tris(3,5-dimethylphenyl carbamate) (Chiralcel® OD). The twoenantiomers, (R)-20 or (S)-20, are separated with a resolution greaterthan 3.0 by using a mobile phase made of 10% ethanol and 90% hexanes atroom temperature with a flow rate of 1 mL/min. The UV detectionwavelength is 220 nm. The retention times for (S)-enantiomer ((S)-20)and (R)-enantiomer ((R)-20) are 10.3 minutes (the first peak) and 13.1minutes (the second peak), respectively.

For (R)-20 or (S)-20: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.83 (s, 1H), 8.75(s, 1H), 8.39 (s, 1H), 7.77 (d, 1H, J=3.7 Hz), 7.09 (d, 1H, J=3.7 Hz),5.63 (s, 2H), 4.53 (td, 1H, J=19.4, 4.0 Hz), 3.51 (t, 2H, J=8.1 Hz),3.23 (dq, 2H, J=9.3, 4.3 Hz), 2.41 (m, 1H), 1.79 (m, 1H), 1.66-1.13 (m,7H), 0.81 (t, 2H, J=8.2 Hz), 0.124 (s, 9H); C₂₃H₃₂N₆OSi (MW, 436.63),LCMS (EI) m/e 437 (M⁺+H) and 459 (M⁺+Na).

The following table summarizes analyses and reaction conditions for thisasymmetric hydrogenation.

Metal/Ligand/ H₂ Conversion Major Catalyst Temp. Pressure Time (HPLC %Enantiomer Precursor Solvent (° C.) (Bar) (h) area %) ee (R)- or (S)-20[Ru(p-cymene) MeOH 50 60 69 12 72.7 (S)-20 (S—C3-TunePhos)Cl]Cl (1^(st)peak) [Ru(p-cymene) EtOAc 75 60 19 93 38.9 (S)-20 (S—C3-TunePhos)Cl]Cl(1^(st) peak) [Ru(p-cymene) THF 75 60 19 94 29.9 (S)-20(S—C3-TunePhos)Cl]Cl (1^(st) peak) [Ru(p-cymene) CH₂Cl₂ 75 60 19 99 34.1(S)-20 (S—C3-TunePhos)Cl]Cl (1^(st) peak) [Ru(p-cymene) CH₂Cl₂ 75 60 2197 32.7 (S)-20 (S—C1-TunePhos)Cl]Cl (1^(st) peak) [Ru(p-cymene) CH₂Cl₂75 60 21 97 26.0 (S)-20 (S—C2-TunePhos)Cl]Cl (1^(st) peak) [Ru(p-cymene)CH₂Cl₂ 75 60 21 99 17.4 (S)-20 (S—C4-TunePhos)Cl]Cl (1^(st) peak)[Ru(p-cymene) CH₂Cl₂ 75 60 21 98 7.4 (S)-20 (S—C5-TunePhos)Cl]Cl (1^(st)peak) [Ru(p-cymene) CH₂Cl₂ 75 60 21 91 3.4 (S)-20 (S—C6-TunePhos)Cl]Cl(1^(st) peak)

Structures of chiral phosphine ligands used in this study are listedbelow.

Representative preparative asymmetric hydrogenation procedure isdescribed below.

(S)-3-Cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((S)-20)

A solution of(Z)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylonitrile(19, 116 mg) in methylene chloride (CH₂Cl₂, 4.0 mL) in a pressure glasstube was treated with the catalyst [Ru(p-cymene)(S—C3-TunePhos)Cl]Cl(8.5 mg) under nitrogen before the reaction mixture was pressurized withhydrogen gas to 60 bar pressure. The reaction mixture was stirred at 75°C. under this hydrogen pressure for 19 h. When HPLC analysis showed thatthe substrate was completely consumed, the reaction mixture was cooleddown to room temperature. The enantiomeric excess of the reactionmixture was determined to be 34.1% ee (67.05% of the first peak, (S)-20;32.95% of the second peak, (R)-20) by chiral HPLC analysis. The reactionmixture was then filtered through a thin silica gel pad and the pad waswashed with methylene chloride (CH₂Cl₂, 5 mL). The filtrate was thenconcentrated under reduced pressure to dryness. The resultant foamysolid (107 mg) was analyzed by chiral HPLC analysis and result showed a34.1% of enantiomeric excess favoring the first peak (67.05% of thefirst peak, (S)-20; 32.95% of the second peak, (R)-20).

(E)-Methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylate(21)

To a stirred suspension of4-(1H-pyrazol-4-yl)-7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidine(17, 12.08 g, 38.31 mmol) and methyl 3-cyclopentylprop-2-ynoate (18,8.970 g, 45.97 mmol, 1.2 equiv) in acetonitrile (76 mL, 1400 mmol) atroom temperature was added 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 2.92mL, 19.2 mmol, 0.5 equiv). The resulting reaction mixture was stirred atroom temperature for 2 h. When LCMS showed the reaction was deemedcomplete, the reaction mixture was quenched with water (50 mL) and 1 Naqueous HCl solution (20 mL). The quenched reaction mixture was adjustedto pH 4 after treatment with 1 N aqueous HCl solution. The mixture wasthen extracted with EtOAc (2×100 mL) and the combined organic layerswere washed with brine, dried over magnesium sulfate (MgSO₄), filteredand concentrated under reduced pressure. The residue was purified byCombiflash (SiO₂, 0-50% EtOAc/hexane gradient elution) to afford(E)-methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylate(21, 6.838 g, 17.92 g theoretical, 38% in yield) as a colorless, veryviscous oil. For 19: ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.93 (s, 1H), 8.55(bs, 1H), 8.44 (s, 1H), 7.49 (d, 1H, J=3.5 Hz), 6.86 (d, 1H, J=3.5 Hz),6.34 (s, 1H), 5.74 (s, 2H), 4.56 (m, 1H), 3.84 (s, 3H), 3.60 (t, 2H,J=8.2 Hz), 2.01 (m, 2H), 1.96 (m, 4H), 1.77 (m, 2H), 0.98 (t, 2H, J=8.2Hz), 0.00 (s, 9H); C₂₄H₃₃N₅O₃Si (MW, 467.64), LCMS (EI) m/e 468.2(M⁺+H).

(R)-Methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((R)-22) and (S)-Methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((S)-22)

General screening procedure for asymmetric hydrogenation using thesubstrate, (E)-methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylate(21), to afford optically enriched product, methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((R)-22 or (S)-22): A 300 mL-volume autoclave with glass vial (20 mL)was charged with the substrate (21), the catalyst (metal, ligand, andcatalyst precursor), and oxygen-free solvent (4-6 mL) under nitrogen.This autoclave was charged with hydrogen gas to the desired pressure andstirred at room temperature or heated with oil bath. After hydrogen gaswas released, the reaction mixture was concentrated under reducedpressure. The residue was purified by eluting through a silica gel padusing a mixture of ethyl acetate and methanol (v/v=9/1) to affordproduct, methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((R)-22 or (S)-22), for chemical conversion (HPLC and chiral HPLC),LC/MS and NMR spectroscopy and enantiomeric excess (% ee by chiral HPLC)determination.

The determination of enantiomeric excess (% ee) of the product wascarried out by chiral HPLC analysis. A chiral HPLC method was developedusing a Chiralcel®OD-H column (4.6×250 mm, 5 μm), purchased from ChiralTechnologies, Inc., packed with a silicagel coated with cellulosetris(3,5-dimethylphenyl carbamate) (Chiralcel® OD). The two enantiomers(R)-22 or (S)-22, are separated with a resolution greater than 3.0 byusing a mobile phase made of 15% ethanol and 85% hexanes at roomtemperature with a flow rate of 1 mL/min. The UV detection wavelength is254 nm. The retention times for (S)-enantiomer ((S)-22) and(R)-enantiomer ((R)-22) are 5.3 minutes (the first peak) and 8.2 minutes(the second peak), respectively.

For (R)-22 or (S)-22: C₂₄H₃₅N₅O₃Si (MW, 469.65), LCMS (EI) m/e 470(M⁺+H) and 492 (M⁺+Na).

The following table summarizes analyses and reaction conditions for thisasymmetric hydrogenation.

Metal/Ligand/ H₂ Conversion Major Catalyst Temp. Pressure Time (HPLCEnantiomer Precursor Solvent (° C.) (Bar) (h) area %) % ee (R)- or(S)-22 Rh(COD)(SSRR-TangPhos)(BF₄) CH₂Cl₂ 50 60 17 99 93.1 (S)-22(1^(st) peak) Rh(COD)(SSRR-TangPhos)(BF₄) MeOH 15 60 67 99 92.7 (S)-22(1^(st) peak) Rh(COD)(SSRR-TangPhos)(BF₄) EtOAc 15 60 67 99 89.7 (S)-22(1^(st) peak) Rh(COD)(SSRR-TangPhos)(BF₄) THF 15 60 67 99 90.1 (S)-22(1^(st) peak) Rh(COD)(+)- DuanPhos)(BF₄) CH₂Cl₂ 15 60 67 99 95.9 (S)-22(1^(st) peak) Rh(COD)(+)- DuanPhos)(BF₄) MeOH 15 60 67 99 92.3 (S)-22(1^(st) peak) Rh(COD)(+)- DuanPhos)(BF₄) EtOAc 15 20 19 99 97.9 (S)-22(1^(st) peak) Rh(COD)(+)- DuanPhos)(BF₄) THF 15 20 19 99 97.0 (S)-22(1^(st) peak) Rh(COD)(−)- DuanPhos)(BF₄) EtOAc 35 20 21 25 95.1 (R)-22(2^(nd) peak) Rh(COD)(−)- DuanPhos)(BF₄) THF 35 50 22 73 94.7 (R)-22(2^(nd) peak)

Structures of chiral phosphine ligands used in this study are listedbelow.

Representative preparative asymmetric hydrogenation procedures aredescribed below.

(S)-Methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((S)-22)

A solution of (E)-methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylate(21, 109 mg) in ethyl acetate (EtOAc, 5.0 mL) in a pressure glass tubewas treated with the catalyst [Rh(COD)(+)-DuanPhos](BF₄) (5.5 mg) undernitrogen before the reaction mixture was pressurized with hydrogen gasto 20 bar pressure. The reaction mixture was stirred at room temperatureunder this hydrogen pressure for 19 h. When HPLC analysis showed thatthe substrate was completely consumed, the reaction mixture was cooleddown to room temperature. The enantiomeric excess of the reactionmixture was determined to be 97.9% ee (98.95% of the first peak, (S)-22;1.05% of the second peak, (R)-22) by chiral HPLC analysis. The reactionmixture was then filtered through a thin silica gel pad and the pad waswashed with ethyl acetate (EtOAc, 5 mL). The filtrate was thenconcentrated under reduced pressure to dryness. The resultant foamysolid (98 mg) was analyzed by chiral HPLC analysis and result showed a97.9% of enantiomeric excess favoring the first peak (98.95% of thefirst peak, (S)-22; 1.05% of the second peak, (R)-22).

(R)-Methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((R)-22)

A solution of (E)-methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)acrylate(21, 815 mg) in tetrahydrofuran (THF, 8.0 mL) in a pressure glass tubewas treated with the catalyst [Rh(COD)(−)-DuanPhos](BF₄) (4.6 mg) undernitrogen before the reaction mixture was pressurized with hydrogen gasto 50 bar pressure. The reaction mixture was stirred at 35° C. underthis hydrogen pressure for 22 h. When HPLC analysis showed that thesubstrate was almost completely consumed, the reaction mixture wascooled down to room temperature. The enantiomeric excess of the reactionmixture was determined to be 94.7% ee (97.35% of the second peak,(R)-22; 2.65% of the first peak, (S)-22) by chiral HPLC analysis. Thereaction mixture was then filtered through a thin silica gel pad and thepad was washed with tetrahydrofuran (THF, 5 mL). The filtrate was thenconcentrated under reduced pressure to dryness. The resultant foamysolid (778 mg) was analyzed by chiral HPLC analysis and result showed a94.7% of enantiomeric excess favoring the second peak (97.35% of thesecond peak, (R)-22; 2.65% of the first peak, (S)-22).

(3R)-3-Cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoicacid ((R)-23)

To a stirred solution of (3R)-methyl3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoate((R)-22, 2.47 g, 5.26 mmol) in THF (30 mL) at room temperature was addeda solution of lithium hydroxide monohydrate (LiOH—H₂O, 265 mg, 6.31mmol, 1.2 equiv) in water (15 mL). The reaction mixture was stirred atroom temperature for 3 h. When LCMS showed the reaction was complete,the reaction mixture was then acidified with 1 N aqueous HCl solution topH 5 before it was extracted with EtOAc (2×25 mL). The combined organiclayers were washed with brine, dried over magnesium sulfate (MgSO₄),filtered and concentrated under reduced pressure to afford(3R)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoicacid ((R)-23, 2.40 g, 2.40 g theoretical, 100% yield) as a colorlessoil, which solidified upon standing at room temperature in vacuo. For(R)-23: ¹H NMR (CDCl₃, 300 MHz) δ ppm 8.95 (s, 1H), 8.95 (bs, 1H), 8.36(s, 1H), 7.57 (d, 1H, J=3.7 Hz), 6.99 (d, 1H, J=3.7 Hz), 5.74 (s, 2H),4.65 (dt, 1H, J=3.1, 10.3 Hz), 3.58 (t, 2H, J=8.2 Hz), 3.24 (dd, 1H,J=16.5, 10.3 Hz), 3.04 (dd, 1H, J=16.2, 3.1 Hz), 2.59 (m, 1H), 2.00 (m,1H), 1.77-1.24 (m, 7H), 0.97 (t, 2H, J=8.2 Hz), 0.00 (s, 9H);C₂₃H₃₃N₅O₃Si (MW, 455.63), LCMS (EI) m/e 456.1 (M⁺+H).

(3R)-3-Cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanamide((R)-24)

To a stirred solution of(3R)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanoicacid ((R)-23, 20 mg, 0.044 mmol) in DMF (1 mL) at room temperature wasadded N,N-carbonyldiimidazole (CDI, 21 mg, 0.13 mmol, 3.0 equiv). Thereaction mixture was then stirred at room temperature and TLC was usedto follow the reaction for formation of acyl imidazole (consumption ofacid to a higher Rf spot with 30% EtOAc/hexane). When TLC showed thatthe acyl imidazole transformation was complete, ammonia gas was thenbubbled through the stirred solution for 30 min to afford the amide(followed by LCMS). The excess amount of ammonia gas was evaporated bybubbling nitrogen vigorously through the solution. The crude product,(3R)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanamide((R)-24), in DMF was used directly to the following reaction to convertamide ((R)-24) into the corresponding nitrile ((R)-20).

(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-20)

Method A. To a stirred solution of(3R)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanamide((R)-24, 20 mg, 0.044 mmol) in DMF (1 mL) at 0° C. was added methylenechloride (1 mL) and triethylamine (0.12 mL, 0.88 mmol, 20.0 equiv),followed by trichloroacetyl chloride (0.052 ml, 0.462 mmol, 10.5 equiv).The resulting reaction mixture was stirred at 0° C. for 1 h. When LCMSshowed the reaction was complete, the reaction mixture was quenched withsaturated sodium bicarbonate solution (NaHCO₃, 5 mL) before beingextracted with EtOAc (2×10 mL). The combined organic layers were washedwith brine, dried over magnesium sulfate (MgSO₄), filtered andconcentrated under reduced pressure. The residue was purified by silicagel chromatography with 0-75% EtOAc/hexane gradient elution to give(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-20, 10 mg, 19 mg theoretical, 53% yield). For (R)-20: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.83 (s, 1H), 8.75 (s, 1H), 8.39 (s, 1H), 7.77(d, 1H, J=3.7 Hz), 7.09 (d, 1H, J=3.7 Hz), 5.63 (s, 2H), 4.53 (td, 1H,J=19.4, 4.0 Hz), 3.51 (t, 2H, J=8.1 Hz), 3.23 (dq, 2H, J=9.3, 4.3 Hz),2.41 (m, 1H), 1.79 (m, 1H), 1.66-1.13 (m, 7H), 0.81 (t, 2H, J=8.2 Hz),0.124 (s, 9H); C₂₃H₃₂N₆OSi (MW, 436.63), LCMS (EI) m/e 437 (M⁺+H) and459 (M⁺+Na).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base)

Method B. To a solution of(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-20, 463 g, 1.06 mol, 98.6% ee) in acetonitrile (4.5 L) was addedwater (400 mL) followed immediately by lithium tetrafluoroborate (LiBF₄,987.9 g, 10.5 mol, 10.0 equiv) at room temperature. The reactiontemperature was observed to decrease from ambient to 12° C. uponaddition of the water and then increase to 33° C. during the addition oflithium tetrafluoroborate (LiBF₄). The resulting reaction mixture washeated to reflux (about 80° C.) for overnight. An aliquot was quenchedinto ethyl acetate/water and checked by LCMS and TLC (95:5 ethylacetate/methanol, v/v). When LCMS and TLC analyses showed both thehydroxylmethyl intermediate ((R)-25) and fully de-protected material((R)-13, free base) produced but no starting material ((R)-20) left, thereaction mixture was cooled gradually to <5° C. before a 20% aqueoussolution of ammonium hydroxide (NH₄OH, 450 mL) was added gradually toadjust the pH of the reaction mixture to 9 (checked with pH strips). Thecold bath was removed and the reaction mixture was gradually warmed toroom temperature and stirred at room temperature for overnight. Analiquot was quenched into ethyl acetate/water and checked by LCMS andTLC (95:5 ethyl acetate/methanol, v/v) to confirm completede-protection. When LCMS and TLC showed the reaction was deemedcomplete, the reaction mixture was filtered and the solids were washedwith acetonitrile (1 L). The combined filtrates were then concentratedunder reduce pressure, and the residue was partitioned between ethylacetate (EtOAc, 6 L) and half-saturated brine (3 L). The two layers wereseparated and the aqueous layer was extracted with ethyl acetate (2 L).The combined organic layers were washed with half-saturated sodiumbicarbonate (NaHCO₃, 3 L) and brine (3 L), dried over sodium sulfate(Na₂SO₄), and concentrated under reduced pressure to give the crudeproduct as an orange oil. The crude material was then purified by flashcolumn chromatography (SiO₂, 40 to 100% ethyl acetate/heptane gradientelution) to afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base, 273 g, 324.9 g theoretical, 84% yield) as a whitefoam. This material was checked by ¹⁹F NMR to ensure no lithiumtetrafluoroborate (LiBF₄) remained and by chiral HPLC (Chiralcel® OD,90:10 hexane/ethanol) to confirm enantiomeric purity and was usedwithout further purification to prepare the corresponding phosphatesalt. For (R)-13 (free base): ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 12.1 (bs,1H), 8.80 (d, 1H, J=0.42 Hz), 8.67 (s, 1H), 8.37 (s, 1H), 7.59 (dd, 1H,J=2.34, 3.51 Hz), 6.98 (dd, 1H, J=1.40, 3.44 Hz), 4.53 (td, 1H, J=19.5,4.63 Hz), 3.26 (dd, 1H, J=9.77, 17.2 Hz), 3.18 (dd, 1H, J=4.32, 17.3Hz), 2.40 (m, 1H), 1.79 (m, 1H), 1.65 to 1.13 (m, 7H); C₁₇H₁₈N₆(MW,306.37) LCMS (EI) m/e 307 (M⁺+H).

(2E)-3-Cyclopentylacrylaldehyde (28)

To a stirred suspension of triphenylphosphoranylidene)acetaldehyde (27,62.75 g, 200.0 mmol, 1.0 equiv) in anhydrous benzene (400 mL, 4476 mmol)was added cyclopentanecarbaldehyde (26, 21.36 mL, 200.0 mmol) at roomtemperature. The resulting reaction mixture was then heated at 80° C.for 16 h. When TLC and HPLC showed that the reaction was deemedcomplete, the reaction mixture was concentrated under reduced pressure.The residue was then directly purified by Combiflash (SiO₂) with 0-10%EtOAc/hexane gradient elution to afford (2E)-3-cyclopentylacrylaldehyde(28, 14.4 g, 24.84 g theoretical, 58% yield) as a yellow oil. For 28: ¹HNMR (DMSO-d₆, 400 MHz) δ ppm ¹H NMR (CDCl₃, 400 MHz) δ 9.49 (d, 1H,J=7.8 Hz), 6.82 (dd, 1H, J=15.6, 7.8 Hz), 6.08 (dd, 1H, J=15.6, 8.0 Hz),2.72 (m, 1H), 1.89 (m, 2H), 1.67 (m, 4H), 1.44 (m, 2H); C₈H₁₂O (MW,124.18) LCMS (EI) m/e 125 (M⁺+H).

(2R)-1-Ethyl 2-methylpyrrolidine-1,2-dicarboxylate ((R)-30)

To a stirred suspension of D-proline ((R)-29, 13.955 g, 120.0 mmol) andpotassium carbonate (K₂CO₃, 33.17 g, 240.0 mmol, 2.0 equiv) in anhydrousmethanol (MeOH, 240 mL, 5925 mmol) at 0° C. was added ethylchloroformate (28.4 mL, 288 mmol, 2.4 equiv) at room temperature. Theresulting reaction mixture was then stirred at room temperature for 18h. When LCMS showed the reaction was deemed complete, the solvent wasremoved under reduced pressure. The resulting residue was then treatedwith water (80 mL) and saturated aqueous NaHCO₃ (80 mL) before beingextracted with EtOAc (2×100 mL). The combined organic layers were washedwith brine, dried over magnesium sulfate (MgSO₄), filtered, andconcentrated under reduced pressure to give the pure (2R)-1-ethyl2-methylpyrrolidine-1,2-dicarboxylate ((R)-30, 18.792 g, 24.14 gtheoretical, 77.8% yield) as a colorless volatile oil. For (R)-30: ¹HNMR (CDCl₃, 400 MHz) δ ppm 4.35 (dd, 0.5H, J=8.7, 3.5 Hz), 4.28 (dd,0.5H, J=8.7, 3.7 Hz), 4.13 (m, 2H), 3.72 (s, 1.5H), 3.70 (s, 1.5H),3.59-3.41 (m, 2H), 2.20 (m, 1H), 2.01-1.86 (m, 3H), 1.25 (t, 1.5H, J=7.1Hz), 1.18 (t, 1.5H, J=7.1 Hz); C₉H₁₅NO₄(MW, 201.22), LCMS (EI) m/e 201.9(M⁺+H).

(7aR)-1,1-Bis(3,5-bis(trifluoromethyl)phenyl)tetrahydropyrrolo[1,2-c]oxazol-3(1H)-one((R)-32)

To a stirred solution of 3,5-bis(trifluoromethyl)bromobenzene (31, 15.2mL, 60.0 mmol, 3.0 equiv) in anhydrous THF (50 mL) at 0° C. was added asolution of 2.0 M of isopropylmagnesium chloride (iPrMgCl) intetrahydrofuran (THF, 31.5 mL) dropwise. The resulting mixture wasstirred at 0° C. for 1 h before being treated with a solution of(2R)-1-ethyl 2-methylpyrrolidine-1,2-dicarboxylate ((R)-30, 4.024 g,20.0 mmol) in anhydrous THF (14 mL) drop wise at 0° C. After theaddition, the ice bath was removed and the reaction mixture was heatedto 65° C. and stirred at 65° C. for 5 h. When LCMS showed that thereaction was deemed complete, the reaction mixture was quenched withsaturated aqueous NH₄Cl solution (120 mL) and extracted with EtOAc(2×100 mL). The combined organic layers were washed with brine, driedover magnesium sulfate (MgSO₄), filtered and concentrated under reducedpressure to afford the crude(7aR)-1,1-bis(3,5-bis(trifluoromethyl)phenyl)tetrahydropyrrolo[1,2-c]oxazol-3(1H)-one((R)-32, 11.03 g, 100%) as a viscous oil, which was directly used in thesubsequent reaction without further purification. For crude (R)-32:C₂₂H₁₃F₁₂NO₂ (MW, 551.32), LCMS (EI) m/e 552 (M⁺+H).

(2R)-Bis(3,5-bis(trifluoromethyl)phenyl)(pyrrolidin-2-yl)methanol((R)-33)

To a stirred solution of crude(7aR)-1,1-bis(3,5-bis(trifluoromethyl)phenyl)tetrahydropyrrolo[1,2-c]oxazol-3(1H)-one((R)-32, 11.03 g, 20.0 mmol) in methanol (MeOH, 80 mL, 1975 mmol) wasadded solid potassium hydroxide (KOH, 3.366 g, 60.0 mmol, 3.0 equiv) atroom temperature. The resulting dark reaction mixture was heated to 65°C. and stirred at 65° C. for 22 h. When LCMS showed the reaction wasdeemed complete, the reaction mixture was cooled to room temperaturebefore the solvent was evaporated under reduced pressure. The residuewas then treated with water (100 mL) and extracted with EtOAc (2×100mL). The combined organic layers were washed with brine, dried overmagnesium sulfate (MgSO₄), filtered, and concentrated under reducedpressure. The residue was then purified by Combiflash (SiO₂) with 0-30%EtOAc/hexane gradient elution to afford(2R)-bis(3,5-bis(trifluoromethyl)phenyl)(pyrrolidin-2-yl)methanol((R)-33, 8.30 g, 10.51 g theoretical, 79% yield for 2 steps) as a yellowviscous paste. For (R)-33: ¹H NMR (CD₃OD, 400 MHz) δ ppm 8.24 (s, 2H),8.16 (s, 2H), 7.85 (s, 2H), 4.49 (t, 1H, J=7.7 Hz), 2.92 (m, 2H), 1.74(m, 2H), 1.67 (m, 1H), 1.55 (m, 1H); C₂₁H₁₅F₁₂NO (MW, 525.33), LCMS (EI)m/e 526.0 (M⁺+H).

(2R)-2-Bis[3,5-bis(trifluoromethyl)-phenyl][(trimethylsilyl)oxy]-methylpyrrolidine((R)-34)

To a stirred solution of(2R)-bis(3,5-bis(trifluoromethyl)phenyl)(pyrrolidin-2-yl)methanol((R)-33, 8.30 g, 14.2 mmol) and triethylamine (TEA, 5.98 mL, 42.6 mmol,3.0 equiv) in anhydrous methylene chloride (CH₂Cl₂, 56.0 mL, 874 mmol)at 0° C. was added trimethylsilyl trifluoromethanesulfonate (TMSOTf,3.89 mL, 21.3 mmol, 1.5 equiv). The resulting reaction mixture wasstirred at 0° C. for 1 h. When LCMS showed the reaction was deemedcomplete, the reaction mixture was quenched with water (80 mL) andextracted with EtOAc (2×100 mL). The combined organic layers were washedwith brine, dried over magnesium sulfate (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified byCombiflash (SiO₂) with 0-10% EtOAc/hexane gradient elution to give(2R)-2-bis[3,5-bis(trifluoromethyl)phenyl][(trimethylsilyl)oxy]methylpyrrolidine((R)-34, 6.869 g, 8.48 g theoretical, 81% yield) as a very viscousyellow syrup. For (R)-34: ¹H NMR (CDCl₃, 300 MHz) δ ppm 8.08 (s, 2H),7.92 (s, 2H), 7.84 (s, 2H), 4.32 (t, 1H, J=7.2 Hz), 2.98 (m, 1H), 2.63(m, 1H), 1.79 (m, 1H), 1.58 (m, 2H), 1.20 (m, 1H), 0.00 (s, 9H);C₂₄H₂₃F₁₂NOSi (MW, 597.51), LCMS (EI) m/e 598.0 (M⁺+H).

(2R)-2-Bis[3,5-bis(trifluoromethyl)phenyl][(triethylsilyl)oxy]-methylpyrrolidine((R)-35)

To a stirred solution of(2R)-bis(3,5-bis(trifluoromethyl)phenyl)(pyrrolidin-2-yl)methanol((R)-33, 3.832 g, 7.294 mmol) and 2,6-lutidine (4.27 mL, 36.5 mmol, 5.0equiv) in anhydrous methylene chloride (CH₂Cl₂, 15.0 mL, 234 mmol) at 0°C. was added triethylsilyl trifluoromethanesulfonate (TESOTf, 5.0 mL,21.9 mmol, 3.0 equiv). The resulting reaction mixture was stirred atroom temperature for 21 h. When LCMS showed the reaction was deemedcomplete, the reaction mixture was quenched with saturated aqueousNaHCO₃ solution (70 mL), extracted with EtOAc (2×50 mL). The combinedorganic layers were washed with brine, dried over magnesium sulfate(MgSO₄), filtered, and concentrated under reduced pressure. The residuewas purified by Combiflash (SiO₂) with 0-10% EtOAc/hexane gradientelution to give(2R)-2-bis[3,5-bis(trifluoromethyl)phenyl][(triethylsilyl)oxy]methylpyrrolidine((R)-35, 4.575 g, 4.665 g theoretical, 98% yield) as a very viscouscolorless syrup. For (R)-35: ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.06 (s, 2H),7.86 (s, 2H), 7.76 (s, 2H), 4.29 (m, 1H), 2.94 (m, 1H), 2.53 (m, 1H),1.83 (m, 2H), 1.53 (m, 2H), 0.85 (t, 9H, J=7.8 Hz), 0.34 (q, 6H, J=7.8Hz); C₂₇H₂₉F₁₂NOSi (MW, 639.59), LCMS (EI) m/e 640.0 (M⁺+H).

(2R)-2-(Bis[3,5-bis(trifluoromethyl)phenyl][tert-butyl(dimethyl)silyl]-oxymethyl)-pyrrolidine((R)-36)

To a stirred solution of(2R)-bis(3,5-bis(trifluoromethyl)phenyl)(pyrrolidin-2-yl)methanol((R)-33, 1.051 g, 2.0 mmol) and triethylamine (TEA, 1.68 mL, 12.0 mmol,6.0 equiv) in anhydrous methylene chloride (5.0 mL, 78 mmol) at 0° C.was added tert-butyldimethylsilyl trifluoromethanesulfonate (TBDMSOTf,1.41 mL, 6.0 mmol, 3.0 equiv). The resulting reaction mixture wasstirred at room temperature for 20 h before being heated at 100° C. for10-20 h. When LCMS showed the reaction was deemed complete, the reactionmixture was quenched with water (30 mL) and extracted with EtOAc (2×50mL). The combined organic layers were washed with brine, dried overmagnesium sulfate (MgSO₄), filtered and concentrated under reducedpressure. The residue was purified by Combiflash (SiO₂) with 0-10%EtOAc/hexane gradient elution to give(2R)-2-(bis[3,5-bis(trifluoromethyl)phenyl][tert-butyl(dimethyl)silyl]oxymethyl)pyrrolidine((R)-36, 1.167 g, 1.279 g theoretical, 91.2% yield) as a very viscouscolorless syrup. For (R)-36: ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.09 (s, 2H),7.87 (s, 2H), 7.75 (s, 2H), 4.33 (m, 1H), 2.98 (m, 1H), 2.54 (m, 1H),1.86 (m, 1H), 1.70 (m, 1H), 1.56 (m, 2H), 0.95 (s, 9H), −0.21 (s, 3H),−0.45 (s, 3H); C₂₇H₂₉F₁₂NOSi (MW, 639.59), LCMS (D) m/e 640.4 (M⁺+H).

(1R)-(4-(1-(1-Cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-37)

A solution of (2E)-3-cyclopentylacrylaldehyde (28, 345 mg, 2.50 mmol,5.0 equiv),(2R)-2-bis[3,5-bis(trifluoromethyl)phenyl][(triethylsilyl)oxy]methylpyrrolidine((R)-35, 16 mg, 0.025 mmol, 0.05 equiv) and 4-nitrobenzoic acid (4.3 mg,0.025 mmol, 0.05 equiv) in anhydrous chloroform (CHCl₃, 2.0 mL, 25 mmol)was stirred at room temperature for 10 min before[4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate (5,0.150 g, 0.50 mmol) was added. The resulting reaction mixture wasstirred at room temperature for 23 h. After LCMS showed that thereaction was deemed complete, the reaction mixture was concentratedunder reduced pressure. The residue was directly purified by Combiflashwith 0-80% EtOAc/hexane gradient elution to afford(1R)-(4-(1-(1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-37, 169 mg, 211.8 mg theoretical, 80% yield) as a paleyellow foam. For (R)-37: C₂₃H₂₉N₅O₃ (MW, 423.51), LCMS (EI) m/e 424(M⁺+H).

(R)-(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-12)

Method B. A solution of(1R)-(4-(1-(1-cyclopentyl-3-oxopropyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-37, 169 mg, 0.399 mmol) in tetrahydrofuran (THF, 1.2 mL,15 mmol) at room temperature was added a 14.3 M solution of ammoniumhydroxide (NH₄OH) in water (1.2 mL), followed by iodine (I₂, 112 mg,0.439 mmol, 1.1 equiv). The resulting reaction mixture was stirred atroom temperature for 25 min. When LCMS showed that the reaction wasdeemed complete, the reaction mixture was quenched with 10% aqueousNa₂S₂O₃ (10 mL) before being extracted with EtOAc (2×15 mL). Thecombined organic layers were washed with brine, dried over magnesiumsulfate (MgSO₄), filtered and concentrated under reduced pressure. Theresidue was purified by Combiflash (SiO₂) with 0-60% EtOAc/hexanegradient elution to afford(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-12, 145.6 mg, 167.8 mg theoretical, 86.8% yield) as acolorless foam.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of (R)-12 and (S)-12 by using a Chiralcel®OD-H column(4.6×250 mm, 5 μm) packed with a silicagel coated with cellulosetris(3,5-dimethylphenyl carbamate) (Chiralcel® OD). (purchased fromChiral Technologies, Inc. The two enantiomers ((R)-12 and (S)-12) areseparated with a resolution greater than 3.5 by using a mobile phasemade from 10% ethanol and 90% hexanes at room temperature with a flowrate of 1 mL/min. The UV detection wavelength is 220 nm. The retentiontimes are 14.1 minutes for (S)-12 (the first peak) and 18.7 minutes for(R)-12 (the second peak), respectively.

For (R)-12: achiral purity (99.3 area % by HPLC detected at 220 nm);chiral purity (94.9 area % by chiral HPLC; 89.8% ee); ¹H NMR (DMSO-d₆,400 MHz) δ ppm 8.84 (s, 1H), 8.78 (s, 1H), 8.39 (s, 1H), 7.74 (d, 1H,J=3.7 Hz,), 7.11 (d, 1H, J=3.8 Hz), 6.23 (s, 2H), 4.53 (ddd, 1H, J=9.9,9.6, 4.2 Hz), 3.26 (dd, 1H, J=17.4, 9.9 Hz), 3.19 (dd, 1H, J=17.2, 4.3Hz), 2.41 (m, 1H), 1.87-1.13 (m, 8H), 1.07 (s, 9H); C₂₃H₂₈N₆O₂ (MW,420.51), LCMS (EI) m/e 421.4 (M⁺+H).

(3R)-3-Cyclopentyl-3-[4-(7-[2-(trimethylsilyl)ethoxy]methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanal((R)-38)

A solution of (2E)-3-cyclopentylacrylaldehyde (28, 327 mg, 2.50 mmol,5.0 equiv),(2R)-2-bis[3,5-bis(trifluoromethyl)phenyl][(triethylsilyl)oxy]methylpyrrolidine((R)-35, 32 mg, 0.050 mmol, 0.10 equiv) and 4-nitrobenzoic acid (8.5 mg,0.050 mmol, 0.10 equiv) in anhydrous toluene (5.0 mL, 47 mmol) wasstirred at room temperature for 10 min before4-(1H-pyrazol-4-yl)-7-[2-(trimethylsilyl)ethoxy]methyl-7H-pyrrolo[2,3-d]pyrimidine(17, 158 mg, 0.50 mmol) was added. The resulting reaction mixture wasstirred at room temperature for 24 h. When LCMS showed that the reactionwas deemed complete, the reaction mixture was concentrated under reducedpressure. The residue was directly purified by Combiflash (SiO₂) with0-70% EtOAc/hexane gradient elution to give(3R)-3-cyclopentyl-3-[4-(7-[2-(trimethylsilyl)ethoxy]methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanal((R)-38, 184.1 mg, 219.8 mg theoretical, 83.8% yield) as a pale yellowviscous oil. For (R)-38: C₂₃H₃₃N₅O₂Si (MW, 439.63), LCMS (EI) m/e 440(M⁺+H).

(3R)-Cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-20)

Method B. To a stirred solution of(3R)-3-cyclopentyl-3-[4-(7-[2-(trimethylsilyl)ethoxy]methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanal((R)-38, 184 mg, 0.418 mmol) in tetrahydrofuran (THF, 1.2 mL, 15 mmol)at room temperature was added a solution of 14.3 M of ammonium hydroxide(NH₄OH) in water (1.2 mL), followed by iodine (I₂, 117 mg, 0.460 mmol,1.1 equiv). The resulting reaction mixture was stirred at roomtemperature for 30 min. When LCMS showed that the reaction was complete,the reaction mixture was quenched with 10% aqueous Na₂S₂O₃ (10 mL)before being extracted with EtOAc (2×15 mL). The combined organic layerswere washed with brine, dried over magnesium sulfate (MgSO₄), filtered,and concentrated under reduced pressure. The residue was then purifiedby Combiflash (SiO₂) with 0-50% EtOAc/hexane gradient elution to give(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-20, 148.9 mg, 182.5 mg theoretical, 81.6% yield) as a colorlessviscous oil.

The determination of enantiomeric excess (% ee) of the product ((R)-20)was carried by chiral HPLC analysis. A chiral HPLC method was developedusing a Chiralcel® OD-H column (4.6×250 mm, 5 μm), purchased from ChiralTechnologies, Inc., packed a silicagel coated with cellulosetris(3,5-dimethylphenyl carbamate) (Chiralcel® OD). The two enantiomers,(R)-20 or (S)-20, are separated with a resolution greater than 3.0 byusing a mobile phase made of 10% ethanol and 90% hexanes at roomtemperature with a flow rate of 1 mL/min. The UV detection wavelength is220 nm. The retention times for (S)-enantiomer ((S)-20) and(R)-enantiomer ((R)-20) are 10.3 minutes (the first peak) and 13.1minutes (the second peak), respectively.

For (R)-20: achiral purity (99.0 area % by HPLC detected at 220 nm);chiral purity (94.4 area % by chiral HPLC; 88.8% ee); ¹H NMR (DMSO-d₆,400 MHz) δ ppm 8.83 (s, 1H), 8.75 (s, 1H), 8.39 (s, 1H), 7.77 (d, 1H,J=3.7 Hz), 7.09 (d, 1H, J=3.7 Hz), 5.63 (s, 2H), 4.53 (td, 1H, J=19.4,4.0 Hz), 3.51 (t, 2H, J=8.1 Hz), 3.23 (dq, 2H, J=9.3, 4.3 Hz), 2.41 (m,1H), 1.79 (m, 1H), 1.66-1.13 (m, 7H), 0.81 (t, 2H, J=8.2 Hz), 0.124 (s,9H); C₂₃H₃₂N₆OSi (MW, 436.63), LCMS (EI) m/e 437 (M⁺+H) and 459 (M⁺+Na).

(3R)-3-(4-Bromo-1H-pyrazol-1-yl)-3-cyclopentylpropanal ((R)-40)

A solution of (2E)-3-cyclopentylacrylaldehyde (28, 654 mg, 5.0 mmol, 5.0equiv),(2R)-2-(bis[3,5-bis(trifluoromethyl)phenyl][tert-butyl(dimethyl)silyl]oxymethyl)pyrrolidine((R)-35, 64 mg, 0.10 mmol, 0.10 equiv) and 4-nitrobenzoic acid (17 mg,0.10 mmol, 0.10 equiv) in anhydrous toluene (4.0 mL, 38 mmol) wasstirred at rt for 10 min, then cooled to 0° C. before4-bromo-1H-pyrazole (39, 148 mg, 1.0 mmol) was then added. The resultingreaction mixture was stirred at 0° C. for 22 h. When LCMS showed thereaction was deemed complete, the reaction mixture was concentratedunder reduced pressure. The residue was directly purified by CombiFlash(SiO₂) with 0-30% EtOAc/hexane gradient elution to give(3R)-3-(4-bromo-1H-pyrazol-1-yl)-3-cyclopentylpropanal ((R)-40, 230.5mg, 271.2 mg theoretical, 85% yield) as a pale yellow viscous oil. For(R)-40: C₁₁H₁₅BrN₂O (MW, 271.15), LCMS (EI) m/e 271/273 (M⁺+H).

(3R)-3-(4-Bromo-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile ((R)-41)

To a stirred solution of(3R)-3-(4-bromo-1H-pyrazol-1-yl)-3-cyclopentylpropanal ((R)-40, 230.5mg, 0.85 mmol) in tetrahydrofuran (THF, 2.4 mL, 29 mmol) at roomtemperature was added a solution of 14.3 M of ammonium hydroxide (NH₄OH)in water (2.4 mL), followed by iodine (I₂, 237 mg, 0.935 mmol, 1.1equiv). The resulting reaction mixture was stirred at room temperaturefor 30 min. When LCMS showed that the reaction was complete, thereaction mixture was quenched with 10% aqueous Na₂S₂O₃ solution (15 mL)and extracted with EtOAc (2×15 mL). The combined organic layers werewashed with brine, dried over magnesium sulfate (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified byCombiflash (SiO₂) with 0-30% EtOAc/hexane gradient elution to give(3R)-3-(4-bromo-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile ((R)-41,180.7 mg, 227.9 mg theoretical, 79.3% yield) as a colorless viscous oil.

The determination of enantiomeric excess (% ee) of the product ((R)-41)was carried by chiral HPLC analysis. A chiral HPLC method was developedusing a Chiralcel® OD-H column (4.6×250 mm, 5 μm), purchased from ChiralTechnologies, Inc., packed a silicagel coated with cellulosetris(3,5-dimethylphenyl carbamate) (Chiralcel® OD). The two enantiomers,(R)-41 or (S)-41, are separated with a resolution greater than 3.0 byusing a mobile phase made of 15% ethanol and 85% hexanes at roomtemperature with a flow rate of 1 mL/min. The UV detection wavelength is220 nm. The retention times for (S)-enantiomer ((S)-41) and(R)-enantiomer ((R)-41) are 12.8 minutes (the first peak) and 16.7minutes (the second peak), respectively.

For (R)-41: achiral purity (99.0 area % by HPLC detected at 220 nm);chiral purity (91.7 area % by chiral HPLC; 83.4% ee); ¹H NMR (CDCl₃, 400MHz) δ ppm 7.52 (s, 2H), 4.10 (m, 1H), 3.02 (dd, 1H, J=17.0, 8.6 Hz),2.86 (dd, 1H, J=17.0, 3.9 Hz), 2.47 (m, 1H), 1.90 (m, 1H), 1.72-1.46 (m,5H), 1.23 (m, 1H), 1.13 (m, 1H); C₁₁H₁₄BrN₃ (MW, 268.15), LCMS (EI) m/e268/270 (M⁺+H).

(3R)-3-Cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile((R)-42)

A degassed mixture of(3R)-3-(4-bromo-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile ((R)-41,363 mg, 1.35 mmol),4,4,5,5,4′,4′,5′,5′-octamethyl-[2,2′]bis[1,3,2]dioxaborolanyl] (366 mg,1.43 mmol, 1.06 equiv), tetrakis(triphenylphosphine)palladium(0)(Pd(PPh₃)₄, 47 mg, 0.041 mmol, 0.03 equiv) and potassium acetate (KOAc,402 mg, 4.06 mmol, 3.0 equiv) in anhydrous 1,4-dioxane (4.0 mL, 51 mmol)was heated at 120° C. via microwave for 1 h. When LCMS showed thereaction was complete, the reaction mixture, which contains the crudedesired product,(3R)-3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile((R)-42), was used directly for the subsequent Suzuki reaction withoutfurther workup. For crude (R)-42: C₁₇H₂₆BN₃O₂ (MW, 315.22), LCMS (EI)m/e 316 (M⁺+H).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base)

Method C. To a stirred solution of the crude(3R)-3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile((R)-42, 427 mg, 1.35 mmol) in 1,4-dioxane (4.0 mL, 51 mmol), a reactionmixture generated as described above, was added4-chloropyrrolo[2,3-d]pyrimidine (1, 0.160 g, 1.04 mmol, 0.77 equiv),tetrakis(triphenylphosphine)palladium(0) ((Pd(PPh₃)₄, 36 mg, 0.031 mmol,0.03 equiv) and a solution of potassium carbonate (K₂CO₃, 432 mg, 3.13mmol, 3.0 equiv) in water (2.0 mL, 110 mmol) at room temperature. Theresulting reaction mixture was degassed three times and refilled withnitrogen each time before being heated at 100° C. for 21 h. When LCMSshowed the reaction was complete, the reaction mixture was quenched withsaturated aqueous NaHCO₃ (10 mL) and extracted with EtOAc (2×25 mL). Thecombined organic layers were washed with brine, dried over magnesiumsulfate (MgSO₄), filtered and concentrated under reduced pressure. Theresidue was purified by Combiflash (SiO₂) eluting with 0-100%EtOAc/hexane gradient elution followed by 0-5% MeOH/EtOAc to afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-13, free base, 204.3 mg, 318.6 mg theoretical, 64% yield for 2steps) as a colorless oil, which solidified upon standing at roomtemperature in vacuum. For (R)-13 (free base): ¹H NMR (DMSO-d₆, 400 MHz)δ ppm 12.1 (bs, 1H), 8.80 (d, 1H, J=0.42 Hz), 8.67 (s, 1H), 8.37 (s,1H), 7.59 (dd, 1H, J=2.34, 3.51 Hz), 6.98 (dd, 1H, J=1.40, 3.44 Hz),4.53 (td, 1H, J=19.5, 4.63 Hz), 3.26 (dd, 1H, J=9.77, 17.2 Hz), 3.18(dd, 1H, J=4.32, 17.3 Hz), 2.40 (m, 1H), 1.79 (m, 1H), 1.65 to 1.13 (m,7H); C₁₇H₁₈N₆ (MW, 306.37) LCMS (EI) m/e 307 (M⁺+H).

4-Chloro-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(3a)

To a flask equipped with a nitrogen inlet, addition funnel, thermowell,and mechanical stirrer was added 4-chloro-7H-pyrrolo[2,3-d]pyrimidine(1, 600 g, 3.91 mol) and dimethylacetimide (9.6 L). The mixture wascooled to −5° C. in an ice/brine bath and sodium hydride (NaH, 60 wt %,174 g, 4.35 mol, 1.1 equiv) was added in portions as a solid. Themixture went to a dark solution during 15 minutes andtrimethylsilylethoxymethyl chloride (2, 763 mL, 4.31 mol, 1.1 equiv) wasadded slowly via an addition funnel at a rate that the temperature didnot exceed 5° C. The reaction was stirred for 30 minutes, determined tobe complete by TLC and HPLC, and water (1 L) was slowly added to quenchthe reaction. The mixture was then diluted with water (12 L) and MTBE (8L). The layers were separated and the aqueous was re-extracted with MTBE(8 L). The combined organic layers were washed with water (2×4 L) andbrine (4 L), dried over sodium sulfate (NaSO₄), and solvents removedunder reduced pressure. The residue was dissolved in heptane (2 L),filtered and loaded onto a silica gel (3.5 kg) column eluting withheptane (˜6 L), 95% heptane/ethyl acetate (˜12 L), 90% heptane/ethylacetate (10 L), and finally 80% heptane/ethyl acetate (10 L). The purefractions were combined and concentrated under reduced pressure to give4-chloro-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(3a, 987 g, 1109.8 g theoretical, 88.9% yield) as a pale yellow oil thatpartially solidified to an oily solid on standing at room temperature.For 3a: ¹H NMR (DMSO-d₆, 300 MHz) δ ppm 8.67 (s, 1H), 7.87 (d, 1H, J=3.8Hz), 6.71 (d, 1H, J=3.6 Hz), 5.63 (s, 2H), 3.50 (t, 2H, J=7.9 Hz), 0.80(t, 2H, J=8.1 Hz), 1.24 (s, 9H); ¹³C NMR (DMSO-d₆, 100 MHz) δ ppm 151.3,150.8, 150.7, 131.5, 116.9, 99.3, 72.9, 65.8, 17.1, −1.48; C₁₂H₁₈ClN₃OSi(MW 283.83), LCMS (EI) m/e 284/286 (M⁺+H).

4-Chloro-7-(diethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine (3b)

To a 1 liter round bottom flask equipped with a stir bar, condenser andnitrogen inlet was charged 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1, 31.0g, 0.202 mol) and triethyl orthoformate (330 ml, 2.00 mol, 10.0 equiv).The reaction mixture was warmed to reflux to generate a clear solution.The reaction was checked after 63 hours by HPLC. When the reaction wasdeemed complete, the reaction mixture was concentrated under reducedpressure. The residue was purified by a silica gel flash columnchromatography eluted with a 20% to 25% ethyl acetate/hexane (v/v)gradient (TLC conditions: 30% ethyl acetate/hexane) to afford4-chloro-7-(diethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine (3b, 48.56 g,51.65 g theoretical, 94% yield) as a light yellow oil. For 3b: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.68 (s, 1H), 7.79 (d, 1H, J=3.8 Hz), 6.75 (s,1H), 6.72 (d, 1H, J=3.8 Hz), 3.68 (dd, 2H, J=9.4, 7.2 Hz), 3.54 (dd, 2H,J=9.4, 7.2 Hz), 1.11 (t, 6H, J=7.2 Hz); C₁₁H₁₄ClN₃O₂ (MW, 255.70), LCMS(EI) m/e 182/184 (M⁺+H for corresponding 7-formylation product of 1) and154/156 (M⁺+H for 1).

tert-Butyl 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (3c)

To a 250 mL round bottom flask equipped with a stir bar and nitrogeninlet was charged 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1, 5.00 g,0.0326 mol), 1,4-dioxane (40 ml, 0.500 mol),1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 24.3 mL, 0.163 mol, 5.0 equiv)and 4-(N,N-dimethyl)aminopyridine (DMAP, 0.80 g, 0.0065 mol, 0.2 equiv).To this solution was added di-tert-butyldicarbonate (BOC₂O, 21.2 g,0.0976 mol, 3.0 equiv) in one portion at room temperature. The resultingreaction solution becomes yellow/orange in color with the evolution ofcarbon dioxide. The reaction was monitored by TLC (80% hexane/ethylacetate) and was complete after stirring at room temperature for about24 hours. The reaction mixture was then diluted with 20% aqueous brinesolution (40 mL) and ethyl acetate (40 mL). The two layers wereseparated, and the aqueous layer was extracted with ethyl acetate (40mL). The combined organic extracts were washed with brine, dried overmagnesium sulfate, and concentrated under reduced pressure to yield thecrude, desired product (3c) as a red to orange oil. Flash columnchromatography purification (SiO₂, 0 to 15% ethyl acetate/hexanegradient elution) afforded pure tert-butyl4-chloro-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate (3c, 6.28 g, 8.27 gtheoretical, 75.9% yield) as off-white solids. For 3c: ¹H NMR (DMSO-d₆,400 MHz) δ ppm 8.79 (s, 1H), 7.94 (d, 1H, J=4.0 Hz), 6.80 (d, 1H, J=4.2Hz), 1.60 (s, 9H); C₁₁H₁₂ClN₃O₂ (MW, 253.68), LCMS (EI) m/e 276/278(M⁺+Na).

4-Chloro-7-(triisopropylsilyl)-7H-pyrrolo[2,3-d]pyrimidine (3d)

To a 250 mL oven dried three-neck round bottom flask equipped with astir bar, condenser, septa, nitrogen inlet and thermocouple was chargedsodium hydride (NaH, 60 wt %, 1.56 g, 0.0391 mol, 1.2 equiv) andanhydrous tetrahydrofuran (THF, 26 mL, 0.320 mol). The mixture waschilled to 0-5° C. To a oven dried 100 mL round bottom flask was charged4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1, 5.00 g, 0.0326 mol) andanhydrous tetrahydrofuran (42 mL, 0.520 mol), and the resulting slurrywas then added portion wise via large bore canula over 15 minutes to thesodium hydride (NaH) suspension in THF. The reaction temperature rose to6.8° C. after the addition of the substrate. The reaction mixture wasstirred at 0-5° C. for 40 minutes before being charged neattriisopropylsilyl chloride (6.6 g, 7.24 mL, 0.0342 mol, 1.05 equiv) viasyringe over 5 minutes. The cooling bath was removed and the reactionmixture was warmed to reflux for 4 hours. The reaction was monitored byTLC (80% hexane/ethyl acetate). When the reaction was deemed complete,the reaction mixture was cooled to room temperature and dilute withethyl acetate (100 mL) and 20% aqueous brine (50 mL). The two layerswere separated and the aqueous layer was extracted with ethyl acetate(100 mL). The combined organic fractions were washed with 1M sodiumbicarbonate (NaHCO₃) aqueous solution (100 mL) and 20% aqueous brine(100 mL), dried over magnesium sulfate (MgSO₄), filtered andconcentrated under reduced pressure. The residue was purified by flashchromatography (SiO₂, 10% ethyl acetate/hexane gradient elution) toafford 4-chloro-7-(triisopropylsilyl)-7H-pyrrolo[2,3-d]pyrimidine (3d,10.0 g, 10.10 g theoretical, 99%) as an amber oil. For 3d: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.61 (s, 1H), 7.67 (d, 1H, J=3.7 Hz), 6.76 (d,1H, J=3.5 Hz), 1.86 (m, 3H), 1.02 (d, 18H, J=7.5 Hz); C₁₅H₂₄ClN₃Si (MW,309.91), LCMS (EI) m/e 310/312 (M⁺+H).

7-[(Benzyloxy)methyl]-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (3e)

To a oven dried 250 mL three-neck round bottom flask equipped with astir bar, thermocouple, septa and nitrogen inlet was charged sodiumhydride (NaH, 60 wt %, 1.56 g, 0.0391 mol, 1.2 equiv) and anhydroustetrahydrofuran (THF, 25.0 mL, 0.308 mol) and the resulting mixture waschilled to 0-5° C. To a 100 ml oven dried round bottom flask was charged4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1, 5.00 g, 0.0326 mol) andanhydrous tetrahydrofuran (50 mL, 0.616 mol), and the resulting slurrywas added portion wise via large bore canula over 20 minutes to thesodium hydride (NaH) suspension in THF. The cooling bath was removedafter the addition is complete and the reaction mixture was stirred atroom temperature for 1 hour. The slurry becomes green in color after itis warmed to 16.5° C. The mixture was cooled to 0-5° C. before neatbenzyl chloromethyl ether (5.28 mL, 0.0342 mol, 1.05 equiv) was chargedover 13 minutes via syringe. The cold bath was removed and the reactionmixture was warmed to room temperature gradually and stirred at roomtemperature for 20 h. The reaction mixture was quenched with 20% aqueousbrine (50 mL) and diluted with ethyl acetate (100 mL) when the reactionwas deemed complete. The two layers were separated, and the aqueouslayer was extracted with ethyl acetate (50 mL). The combined organicfractions were dried over magnesium sulfate, filtered, and concentrateunder reduced pressure. The residue was then purified by flashchromatography (SiO₂, 10% to 15% ethyl acetate/hexane gradient elution)to afford 7-[(benzyloxy)methyl]-4-chloro-7H-pyrrolo[2,3-d]pyrimidine(3e, 6.31 g, 8.92 g theoretical, 70.7%) as a green oil. For 3e: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.69 (s, 1H), 7.90 (d, 1H, J=3.7 Hz), 7.26 (m5H), 6.71 (d, 1H, J=3.7 Hz), 5.75 (s 2H), 4.51 (s, 2H); C₁₄H₁₂ClN₃O (MW,273.72), LCMS (EI) m/e 274/276 (M⁺+H).

(4-Chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl pivalate (3f)

To a oven dried 2 L 4-neck round bottom flask equipped with overheadstirring, septa, thermocouple, 500 mL addition funnel and nitrogen inletwas charged sodium hydride (NaH, 60 wt %, 29.7 g, 0.742 mol, 1.34 equiv)and anhydrous tetrahydrofuran (THF, 400 mL, 5.0 mol) and the resultingmixture was cooled to 0-3° C. To a oven dried 1 L round bottom flask wascharged 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1, 85.0 g, 0.553 mol) andtetrahydrofuran (600 mL, 7.0 mol) resulting in a slurry. This resultingslurry was then portion wise added to the suspension of sodium hydridein THF via large bore canula over 27 minutes at 0-5° C. The resultingsolution was heterogeneous and green in color. Following the addition,the cold bath was removed and the mixture was gradually warmed to roomtemperature and allowed to stir at room temperature for 1 hour beforebeing cooled to 0-5° C. Chloromethyl pivalate (pivaloyloxymethylchloride, POM-Cl, 103 ml, 0.692 mol, 1.25 equiv) was added portion wiseinto the reaction mixture over 25 minutes via syringe with stirring at0-5° C. The addition of chloromethyl pivalate (POM-Cl) was mildlyexothermic and the reaction temperature went to as high as 14° C. Afteraddition of chloromethyl pivalate (POM-Cl), the cooling bath was removedand the reaction mixture was allowed to return to room temperature andstirred at room temperature for overnight. When the reaction was deemedcomplete after about 16 hours, the reaction was quenched with 20%aqueous brine (250 mL) and ethyl acetate (250 mL) producing a slurry.Additional amount of water (250 mL) was added until the mixture becomesa homogeneous solution. The two layers were separated and the aqueouslayer was extracted with ethyl acetate (250 mL). The combined organicfractions were dried over magnesium sulfate (MgSO₄), filtered, andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, 10% to 15% ethyl acetate/hexane gradientelution) to afford the desired product as yellow, crystalline solids(155 g). The combined solids were treated with hexanes (750 mL) and theresulting slurry was warmed to 55° C. to produce a homogeneous solution.The resulting solution was then gradually cooled to room temperature andstirred at room temperature for overnight before being cooled to 0-5° C.for 2 h. The solids were collected by filtration, washed with pre-cooledhexanes (2×30 mL), dried in vacuum to afford4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl pivalate (3f, 134.9 g,148.0 g theoretical, 91% yield) as white solids. For 3f: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.71 (s, 1H), 7.83 (d, 1H, J=3.7 Hz), 6.73 (d,1H, J=3.8 Hz), 6.23 (s 2H), 1.06 (s, 9H); ¹³C NMR (DMSO-d₆, 100 MHz) δppm 176.9, 151.2, 151.1, 151.0, 131.6, 117.1, 99.9, 66.9, 38.3, 26.5;C₁₂H₁₄ClN₃O₂ (MW, 267.71), LCMS (EI) m/e 268/270 (M⁺+H).

3-Cyclopentylacrylonitrile (8)

A solution of diethyl cyanomethylphosphonate (7, 742.5 g, 4.2 mol, 1.1equiv) in dry THF (5.75 L) was stirred under nitrogen on anice-water-methanol bath and a solution of 1 M potassium tert-butoxide inTHF (4 L, 4.0 mol, 1.05 equiv) was added at such a rate as to keep thetemperature below 0° C. After addition of 1 M potassium tert-butoxide inTHF was complete, the stirring was continued on the cold bath for 1 hand a solution of cyclopentanecarbaldehyde (6, 374 g, 3.81 mol) in dryTHF (290 mL) was added at such a rate as to maintain the temperaturebelow 0° C. The cold bath was removed, and the reaction mixture wasgradually warmed to room temperature and stirred at room temperature forovernight. When the reaction was deemed complete, the reaction mixturewas partitioned between methyl tert-butyl ether (MTBE, 14 L), water (10L) and brine (6 L). The two layers were separated, and the combinedorganic phase was washed with brine (6 L). The aqueous phase wasextracted with MTBE (10 L) and washed with brine (6 L). The combinedorganic extracts were concentrated under reduced pressure and theresidue was distilled (65-78° C./6 ton) to afford3-cyclopentylacrylonitrile (8, 437.8 g, 461.7 g theoretical, 94.8%yield) as a colorless oil, which was found to be a mixture of E- andZ-isomer. For 8: ¹H NMR (DMSO-d₆, 400 MHz, for Z-isomer) δ ppm 6.58 (t,1H, J=10.6 Hz), 5.55 (dd, 1H, J=10.8, 0.59 Hz), 2.85 (m, 1H), 1.90-1.46(m, 6H), 1.34 (m, 2H) and (for E-isomer) δ ppm 6.83 (q, 1H, J=8.3 Hz),5.66 (dd, 1H, J=16.5, 1.4 Hz), 2.60 (m, 1H), 1.90-1.46 (m, 6H), 1.34 (m,2H); ¹³C NMR (DMSO-d₆, 100 MHz, for Z-isomer) δ ppm 159.8, 116.6, 97.7,42.3, 32.3, 25.1 and (for E-isomer) δ ppm 160.4, 118.1, 97.9, 43.2,31.5, 24.8; C₈H₁₁N (MW, 121.18), GCMS (EI) m/e 120 (M⁺−H).

4-(1H-Pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5)

Method A. To a flask equipped with a reflux condenser, a nitrogen inlet,mechanical stirrer, and a thermowell was added4-chloro-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(3a, 817 g, 2.88 mol) and dioxane (8 L). To this solution was added4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (4, 728 g,3.75 mol, 1.30 equiv) followed by a solution of potassium carbonate(K₂CO₃, 1196 g, 8.67 mol, 3.0 equiv) in water (4 L). The solution wasdegassed by passing a stream of nitrogen through the solution for 15minutes before being treated withtetrakis(triphenylphosphine)palladium(0) (167 g, 0.145 mol, 0.05 equiv)and the resulting reaction mixture was heated at reflux (about 90° C.)for 2 hours. When the reaction was deemed complete by TLC (1:1heptane/ethyl acetate) and LCMS, the reaction mixture was cooled to roomtemperature, diluted with ethyl acetate (24 L) and water (4 L). The twolayers were separated, and the aqueous layer was extracted with ethylacetate (4 L). The combined organic layers were washed with water (2×2L), brine (2 L), dried over sodium sulfate (Na₂SO₄), and concentratedunder reduced pressure. The residue was suspended in toluene (4 L) andthe solvent was removed under reduced pressure. The residue was finallytriturated with methyl tert-butyl ether (MTBE, 3 L) and the solids werecollected by filtration and washed with MTBE (1 L) to afford4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 581.4 g, 908.5 g theoretical, 64% yield) as white crystallinesolids. For 5: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 13.41 (bs, 1H), 8.74 (s,1H), 8.67 (bs, 1H), 8.35 (bs, 1H), 7.72 (d, 1H, J=3.7 Hz), 7.10 (d, 1H,J=3.7 Hz), 5.61 (s, 2H), 3.51 (t, 2H, J=8.2 Hz), 0.81 (t, 2H, J=8.2 Hz),0.13 (s, 9H); C₁₅H₂₁N₅OSi (MW, 315.45), LCMS (EI) m/e 316 (M⁺+H).

Racemic3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound)

Method A. 3-Cyclopentylacrylonitrile (8, 273.5 g, 2.257 mol, 1.20 equiv)and DBU (28 mL, 0.187 mol, 0.10 equiv) was added to a suspension of4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 591.8 g, 1.876 mol) in acetonitrile (4.7 L) at room temperature. Theresulting reaction mixture was heated to 50-60° C. for 17 hours (a clearsolution developed midway through heating) then to 70-80° C. for 8hours. When LCMS analysis showed the reaction was deemed complete, thereaction mixture was cooled to room temperature. The cooled solution wasthen concentrated under reduced pressure to give the crude product (9)as a thick amber oil. The crude product was dissolved in dichloromethane(DCM) and absorbed onto silica gel then dry-loaded onto a silica column(3 Kg) packed in 33% EtOAc/heptanes. The column was eluted with 33%EtOAc/heptanes (21 L), 50% EtOAc/heptanes (28 L), 60% EtOAc/heptanes (12L) and 75% EtOAc/heptanes (8 L). The fractions containing the desiredproduct (9) were combined and concentrated under reduced pressure togenerate a yellow oil, which was transferred to a 3 L flask with EtOAc.The solvent was removed under reduced pressure and the residual EtOAc byco-evaporating with heptanes. The residue was further dried under highvacuum for overnight to afford racemic3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound, 800 g, 819.1 g theoretical, 97.7%yield) as an extremely viscous yellow oil. For 9: ¹H NMR (DMSO-d₆, 400MHz) δ ppm 8.83 (s, 1H), 8.75 (s, 1H), 8.39 (s, 1H), 7.77 (d, 1H, J=3.7Hz), 7.09 (d, 1H, J=3.7 Hz), 5.63 (s, 2H), 4.53 (td, 1H, J=19.4, 4.0Hz), 3.51 (t, 2H, J=8.1 Hz), 3.23 (dq, 2H, J=9.3, 4.3 Hz), 2.41 (m, 1H),1.79 (m, 1H), 1.66-1.13 (m, 7H), 0.81 (t, 2H, J=8.2 Hz), 0.124 (s, 9H);C₂₃H₃₂N₆OSi (MW, 436.63), LCMS (EI) m/e 437 (M⁺+H) and 459 (M⁺+Na).

(3R)-Cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-10) and(3S)-Cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((S)-10)

A slurry of 1.5 Kg of 20-micron Chiralcel® OD chiral stationary phase(CSP) made by Daicel in 3.0 L of isopropanol (IPA) was packed into aPROCHROM Dynamic Axial Compression Column LC110-1 (11 cm ID×25 cm L;Column Void Vol.: approximate 1.5 L) under 150 bar of packing pressure.The packed column was then installed on a Novasep Hipersep HPLC unit.The column and the Hipersep unit were flushed with methanol (17 L)followed by the mobile phase made of a mixture of isopropanol and hexane(2:8 by volume, 17 L). The feed solution was then prepared by dissolving3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound, 2795 g, 6.4 mol) in the mobile phaseto a concentration of 80 g/L. The feed solution was then sequentiallyinjected into the preparative chiral column for separation. Eachinjection was 120 ml in volume. The chiral column was eluted with themobile phase at a flow rate of 570 mL/min at room temperature. Thecolumn elution was monitored by UV at a wavelength of 330 nm. Underthese conditions a baseline separation of the two enantiomers wasachieved. The retention times were 16.4 minutes (Peak 1, the undesired(S)-enantiomer (S)-10) and 21.0 minutes (Peak 2, the desired(R)-enantiomer (R)-10), respectively. The cycle time for each injectionwas 11 minutes and a total of 317 injections were performed for thisseparation process. Fractions for Peak 1 (the undesired (S)-enantiomer,(S)-10) and Peak 2 (the desired (R)-enantiomer, (R)-10) were collectedseparately from each injection. The collected fractions collected werecontinuously concentrated in the 1-square feet and 2-square feetROTOTHERM evaporator, respectively, at 40° C. under reduced pressure(40-120 bar). The residue from each evaporator was further dried underhigh vacuum to constant weight to afford(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-10, 1307 g, 1397.5 g theoretical, 93.5%) from Peak 2 as a lightyellow oil and(3S)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((S)-10, 1418 g, 1397.5 g theoretical, 101.5%) from Peak 1 as an yellowoil.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of SEM-protected compound ((R)-10 and (S)-10) using aChiralcel® OD-H column (4.6×250 mm, 5 μm), purchased from ChiralTechnologies, Inc., packed with silica gel coated with cellulosetris(3,5-dimethylphenyl carbamate) (Chiralcel® OD). The two enantiomersof SEM-protected compound are separated with a resolution greater than3.0 by using a mobile phase made of 10% ethanol and 90% hexanes at roomtemperature with a flow rate of 1 mL/min. The UV detection wavelength is220 nm. The retention times for (S)-enantiomer ((S)-10) and(R)-enantiomer ((R)-10) are 10.3 minutes and 13.1 minutes, respectively.

The quality of each enantiomer separated by preparative chiral HPLCincluding chemical purity (HPLC area % and wt %), chiral purity (chiralHPLC area %), and residual solvents (IPA and hexane) was analyzed andtheir structures are confirmed by NMRs and LC/MS. For (R)-10: achiralpurity (99.0 area % by HPLC detected at 220 nm; 100.1 wt % by HPLCweight percent assay); chiral purity (99.7 area % by chiral HPLC; 99.4%ee); residual solvents (3.7 wt % for IPA; 0.01 wt % for hexane); ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.83 (s, 1H), 8.75 (s, 1H), 8.39 (s, 1H), 7.77(d, 1H, J=3.7 Hz), 7.09 (d, 1H, J=3.7 Hz), 5.63 (s, 2H), 4.53 (td, 1H,J=19.4, 4.0 Hz), 3.51 (t, 2H, J=8.1 Hz), 3.23 (dq, 2H, J=9.3, 4.3 Hz),2.41 (m, 1H), 1.79 (m, 1H), 1.66-1.13 (m, 7H), 0.81 (t, 2H, J=8.2 Hz),0.124 (s, 9H); C₂₃H₃₂N₆OSi (MW, 436.63), LCMS (EI) m/e 437 (M⁺+H) and459 (M⁺+Na). For (S)-10: achiral purity (99.3 area % by HPLC detected at220 nm; 99.9 wt % by HPLC weight percent assay); chiral purity (99.7area % by chiral HPLC; 99.4% ee); residual solvents (4.0 wt % for IPA;0.01 wt % for hexane); ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.83 (s, 1H),8.75 (s, 1H), 8.39 (s, 1H), 7.77 (d, 1H, J=3.7 Hz), 7.09 (d, 1H, J=3.7Hz), 5.63 (s, 2H), 4.53 (td, 1H, J=19.4, 4.0 Hz), 3.51 (t, 2H, J=8.1Hz), 3.23 (dq, 2H, J=9.3, 4.3 Hz), 2.41 (m, 1H), 1.79 (m, 1H), 1.66-1.13(m, 7H), 0.81 (t, 2H, J=8.2 Hz), 0.124 (s, 9H); C₂₃H₃₂N₆OSi (MW,436.63), LCMS (EI) m/e 437 (M⁺+H) and 459 (M⁺+Na).

(3R)-Cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-10) and(3S)-Cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((S)-10)

The racemic mixture was processed on an SMB unit equipped with 8columns. The separation was performed at various scales using variousconditions presented in the examples below. The purity of eachenantiomer was monitored by a chiral HPLC method using the same mobilephase and the same stationary phase used for the separation to allowrapid determination of the purity. In each case both enantiomers wererecovered as concentrated solutions by evaporation under vacuum, eitherusing a rotary evaporator or falling film evaporators. In examples 1 to3 the desired enantiomer is recovered as the raffinate. In example 4 thedesired enantiomer is recovered as the extract. The chiral purity andyield reported are data measured after the SMB unit has been operatedfor at least 10 to 15 cycles to ensure steady state operations. Variousoperating conditions were tested to ensure high purity and high productyield. In examples 1 to 3, the separation using the same stationaryphase and mobile phase is tested on various SMB units with variouscolumn diameters. In example 4 the SMB is operated at two differentoperating pressures. In example 4, the column configuration was changedfrom the classical <2>/<2>/<2>/<2> to <2>/<2>/<3>1<1> to increase thepurity of the raffinate and increase the throughput by increasing thelength of the SMB Zone III.

Example 1 50 g Scale

Column: Chiralcel® OD

Mobile Phase isopropyl alcohol and n-heptane 20/80 (v/v)

Column length 10 cm

Column ID 10 mm

No of columns 8

Feed concentration 80 g/l

Temperature: 25° C.

Parameters Example 1 Column configuration <2>/<2>/<2>/<2> Recycling flowrate (ml/min) 18 Extract flow rate (ml/min) 7.76 Feed flow rate (ml/min)0.25 Raffinate flow rate (ml/min) 1.4 Eluent flow rate (ml/min) 8.91Switch time (min) 1.52 Desired enantiomer purity 99.15% Desiredenantiomer yield  94.8% Productivity (kg enantiomer/d/kg CSP) 0.41

Example 2 25 kg Scale

Column: Chiralcel® OD

Mobile Phase isopropyl alcohol and n-heptane 20/80 (v/v)

Column length 9.5 cm

Column ID 49 mm

No of columns 8

Feed concentration 80 g/l

Temperature: 25° C.

Parameters Example 2 Column configuration <2>/<2>/<2>/<2> Operatingpressure (bar) 25-28 Recycling flow rate (ml/min) 498.9 Extract flowrate (ml/min) 176.4 Feed flow rate (ml/min) 6.58 Raffinate flow rate(ml/min) 57.8 Eluent flow rate (ml/min) 227.6 Switch time (min) 1.11Desired enantiomer purity 99.3% Desired enantiomer yield   85%Productivity (kg enantiomer/d/kg CSP) 0.43

Example 3 100 kg Scale

Column: Chiralcel® OD

Mobile Phase isopropyl alcohol and n-heptane 20/80 (v/v)

Column length 9.0 cm

Column ID 200 mm

No of columns 8

Feed concentration 53.7 g/l

Temperature: 25° C.

Parameters Example 3 Column configuration <2>/<2>/<2>/<2> Operatingpressure (bar) 35 Recycling flow rate (l/h) 355.0 Extract flow rate(l/h) 124.1 Feed flow rate (l/h) 7.0 Raffinate flow rate (l/h) 114.0Eluent flow rate (l/h) 231.1 Switch time (min) 1.80 Desired enantiomerpurity 99.8% Desired enantiomer yield   92% Productivity (kgenantiomer/d/kg CSP) 0.31

Example 4 100 g Scale

Column: (S,S) Whelk-O® 1

Mobile Phase methyl-tert-butyl ether

Column length 10.0 cm

Column ID 10 mm

No of columns 8

Feed concentration 90 g/l

Parameters Example 4a Example 4b Column configuration <2>/<2>/ <2>/<2>/<2>/<2> <3>/<1> Operating pressure (bar) 27 12 Temperature 23 22Recycling flow rate (ml/min) 22.0 9.0 Extract flow rate (ml/min) 9.6 2.8Feed flow rate (ml/min) 0.5 0.3 Raffinate flow rate (ml/min) 5.9 3.0Eluent flow rate (ml/min) 15 5.5 Switch time (min) 0.70 1.48 Desiredenantiomer purity 99.6% 99.8% Desired enantiomer yield   90%   98%Productivity (kg enantiomer/d/kg CSP) 0.92 0.55

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-12, free base)

Method A. To a solution of(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}proprionitrile((R)-10, 463 g, 1.06 mol, 98.6% ee) in acetonitrile (4.5 L) was addedwater (400 mL) followed immediately by lithium tetrafluoroborate (LiBF₄,987.9 g, 10.5 mol, 10.0 equiv) at room temperature. The reactiontemperature was observed to decrease from ambient to 12° C. uponaddition of the water and then increase to 33° C. during the addition oflithium tetrafluoroborate (LiBF₄). The resulting reaction mixture washeated to reflux (about 80° C.) for overnight. An aliquot was quenchedinto ethyl acetate/water and checked by LCMS and TLC (95:5 ethylacetate/methanol, v/v). When LCMS and TLC analyses showed both thehydroxylmethyl intermediate ((R)-11) and fully de-protected material((R)-12, free base) produced but no starting material ((R)-10) left, thereaction mixture was cooled gradually to <5° C. before a 20% aqueoussolution of ammonium hydroxide (NH₄OH, 450 mL) was added gradually toadjust the pH of the reaction mixture to 9 (checked with pH strips). Thecold bath was removed and the reaction mixture was gradually warmed toroom temperature and stirred at room temperature for overnight. Analiquot was quenched into ethyl acetate/water and checked by LCMS andTLC (95:5 ethyl acetate/methanol, v/v) to confirm completede-protection. When LCMS and TLC showed the reaction was deemedcomplete, the reaction mixture was filtered and the solids were washedwith acetonitrile (1 L). The combined filtrates were then concentratedunder reduce pressure, and the residue was partitioned between ethylacetate (6 L) and half-saturated brine (3 L). The two layers wereseparated and the aqueous layer was extracted with ethyl acetate (2 L).The combined organic layers were washed with half-saturated sodiumbicarbonate (NaHCO₃, 3 L) and brine (3 L), dried over sodium sulfate(Na₂SO₄), and concentrated under reduced pressure to give the crudeproduct as an orange oil. The crude material was then purified by flashcolumn chromatography (SiO₂, 40 to 100% ethyl acetate/heptane gradientelution) to afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-12, free base, 273 g, 324.9 g theoretical, 84% yield) as a whitefoam. This material was checked by ¹⁹F NMR to ensure no lithiumtetrafluoroborate (LiBF₄) remained, and by chiral HPLC (Chiralcel® OD-H,90:10 hexane/ethanol) to confirm enantiomeric purity (98.7% ee), and wasused without further purification to prepare the corresponding phosphatesalt. For (R)-12 (free base): ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 12.1 (bs,1H), 8.80 (d, 1H, J=0.42 Hz), 8.67 (s, 1H), 8.37 (s, 1H), 7.59 (dd, 1H,J=2.34, 3.51 Hz), 6.98 (dd, 1H, J=1.40, 3.44 Hz), 4.53 (td, 1H, J=19.5,4.63 Hz), 3.26 (dd, 1H, J=9.77, 17.2 Hz), 3.18 (dd, 1H, J=4.32, 17.3Hz), 2.40 (m, 1H), 1.79 (m, 1H), 1.65 to 1.13 (m, 7H); C₁₇H₁₈N₆(MW,306.37) LCMS (EI) m/e 307 (M⁺+H).

(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile(R)-10

A solution of(R)-3-cyclopentyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-c]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-10, 75.0 g, 0.172 mol, 98.8% ee) in acetonitrile (600 mL) wascooled to 0-5° C. To the cooled solution was added boron trifluoridediethyl etherate (54.4 mL, 0.429 mol) over 10 minutes while maintainingthe internal reaction temperature below 5° C. Following the addition,the cold bath was removed and the reaction mixture was allowed to warmto room temperature. When HPLC analysis indicated that the level of(R)-10 was below 1%, the initial phase of the deprotection reaction wasconsidered complete. The reaction was then cooled to 0-5° C., followedby the slow addition of water (155 mL). Following the water addition,the cold bath was removed and the resulting reaction mixture was allowedto warm to 13-17° C., and stirred for an additional 2-3 hours. Theresulting reaction mixture was cooled again to 0-5° C. To the cooledreaction mixture was added slowly a solution of ammonia in water[prepared by mixing aqueous 28% ammonia solution (104.5 mL) and water(210.5 mL)] while maintaining the internal reaction temperature at below5° C. After the aqueous ammonia solution was added, the cold bath wasremoved and the reaction was allowed to warm to room temperature. Thehydrolysis was deemed complete when the level of the hydroxylmethylintermediate was below 1% by HPLC analysis.

The resulting reaction mixture was diluted with ethyl acetate (315 mL)and washed with 20% brine (315 mL). The aqueous fraction was backextracted with ethyl acetate (315 mL). The organic fractions werecombined and concentrated under vacuum with a bath temperature of 40° C.to a volume of 380 mL. The concentrated residue was diluted with ethylacetate (600 mL) and washed with 1M NaHCO₃ (2×345 mL) and 20% brine (345mL). The aqueous washes were combined and back extracted with ethylacetate (345 mL). The organic fractions were combined and polishfiltered into a clean 2 L round bottom flask. The organic fraction waswashed with warm water (50° C., 2×450 mL) and then treated withactivated charcoal at 65° C. with stirring for 1.5 hours. The slurry wasfiltered through a celite bed. The filtrate was concentrated undervacuum with a bath temperature of 40° C. The resulting syrup was placedunder high vacuum to provide(R)-3-(4-(7H-pyrrolo[2,3-c]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile[(R)-12, 54.2 g, 103% yield] as a light yellow foam. This material waschecked by ¹⁹F NMR to ensure that the product was not contaminated byany fluorinated impurities. The chemical purity of the isolated freebase was 96.3%. The chiral purity of the free base was 98.8% by chiralHPLC (chiralcel OD, 90:10 hexane/ethanol). The free base was usedwithout further purification to prepare the phosphate salt. ¹H NMR(DMSO-d₆, 400 MHz) δ 12.11 (bs, 1H), 8.79 (d, 1H, J=0.43 Hz), 8.67 (s,1H), 8.37 (s, 1H), 7.59 (q, 1H, J=2.3 Hz), 6.98 (q, 1H, J=1.6 Hz), 4.53(td, 1H, J=19.2, 4.1 Hz), 3.22 (dq, 2H, J=9.8, 4.3 Hz), 2.40 (m, 1H),1.79 (m, 1H), 1.65-1.13 (m, 7H). C₁₇H₁₆N₆ (MW, 306.37), LCMS (EI) m/e307 (M⁺+H).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt ((R)-13, phosphate)

Method A. To a solution of(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-12, free base, 572 g, 1.87 mol) in isopropanol (IPA, 8 L) at 60-65°C. was added a solution of phosphoric acid (186.2 g, 1.9 mol, 1.10equiv) in isopropanol (1.6 L). No exotherm was observed while adding asolution of phosphoric acid, and a precipitate was formed almostimmediately. The resulting mixture was then heated at 76° C. for 1.5hours, then cooled gradually to ambient temperature and stirred at roomtemperature for overnight. The mixture was filtered and the solids werewashed with a mixture of heptanes and isopropanol (1/1, v/v, 3 L) beforebeing transferred back to the original flask and stirred in heptanes (8L) for one hour. The solids were collected by filtration, washed withheptanes (1 L), and dried in a convection oven in vacuum at 40° C. to aconstant weight to afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt ((R)-13, phosphate, 634.2 g, 755 g theoretical, 84%yield) as white to off-white crystalline solids. For (R)-13, phosphate:mp. 197.6° C.; ¹H NMR (DMSO-d₆, 500 MHz) δ ppm 12.10 (s, 1H), 8.78 (s,1H), 8.68 (s, 1H), 8.36 (s 1H), 7.58 (dd, 1H, J=1.9, 3.5 Hz), 6.97 (d,1H, J=3.6 Hz), 4.52 (td, 1H, J=3.9, 9.7 Hz), 3.25 (dd, 1H, J=9.8, 17.2Hz), 3.16 (dd, 1H, J=4.0, 17.0 Hz), 2.41, (m, 1H), 1.79 (m, 1H), 1.59(m, 1H), 1.51 (m, 2H), 1.42 (m, 1H), 1.29 (m, 2H), 1.18 (m, 1H); ¹³C NMR(DMSO-d₆, 125 MHz) δ ppm 152.1, 150.8, 149.8, 139.2, 131.0, 126.8,120.4, 118.1, 112.8, 99.8, 62.5, 44.3, 29.1, 29.0, 24.9, 24.3, 22.5;C₁₇H₁₈N₆ (MW, 306.37 for free base) LCMS (EI) m/e 307 (M⁺+H, base peak),329.1 (M⁺+Na).

Method B. To a solution of(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile((R)-12, 54.2 g, 177 mol) in dichloromethane (782 mL) and 2-propanol(104 mL) at reflux was added a solution of phosphoric acid (19.9 g,0.173 mol, 1.15 equiv) in 2-propanol (34.0 mL) over a period of 47minutes. Following the acid addition, the resulting mixture was heatedto reflux for an additional 1 hour. The mixture was gradually cooled toambient temperature and stirred for 3 hours. The solids were collectedby filtration and washed with dichloromethane (390 mL), followed byn-heptane (390 mL). The solids were partially dried under vacuum at roomtemperature and then under vacuum at 62° C. to afford(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrilephosphate (60.1 g, 84% yield) as white to off-white crystalline solids.Analysis by chiral HPLC (chiralcel OD, 90:10 hexane/ethanol) gave theenantiopurity as 99.2% ee. ¹H NMR (DMSO-d₆, 400 MHz) δ 12.11 (bs, 1H),8.79 (d, 1H, J=0.59 Hz), 8.67 (s, 1H), 8.36 (s, 1H), 7.59 (q, 1H, J=2.3Hz), 6.98 (q, 1H, J=1.6 Hz), 4.53 (td, 1H, J=19.6, 4.4 Hz), 3.22 (dq,2H, J=9.6, 4.3 Hz), 2.40 (m, 1H), 1.79 (m, 1H), 1.65-1.13 (m, 7H).C₁₇H₂₁N₆O₄P (MW, 404.36), LCMS (EI) m/e 307 (M⁺+H) and m/e 329 (M⁺+Na).

(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrilephosphate

Into a 1 L round bottom flask, equipped with stir bar, distillationhead, addition funnel and heating mantle, were charged methanol (520 mL)and(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrilephosphate ((R)-13, phosphate, 40.0 grams, 98.92 mmol). The slurry washeated to 55° C. to generate a slightly pink solution. The solution wascooled to 50° C. and filtered into a 2 L flask equipped with an overheadstirrer, distillation head, addition funnel and heating mantle. The 1 Lround bottom flask and the filter funnel were rinsed with additionalmethanol (104.0 mL). The filtrate solution was heated to reflux todistill methanol (281 mL) over 1 hour under atmospheric pressure.Isopropyl alcohol (IPA) (320 mL) was charged slowly via the additionfunnel over 80 minutes while maintaining the internal temperatureapproximately at 65° C. Precipitation of the phosphate salt was observedduring IPA addition. After the addition of IPA was complete, n-heptane(175 mL) was added slowly at the same temperature. Distillation wascontinued under atmospheric pressure. Additional n-heptane (825 mL) wasadded at approximately the same rate as the distillation rate whilemaintaining the internal temperature at approximately 65° C. Thedistillation was complete when the volume of the distillate reached 742mL (excluding the volume of 281 mL of methanol from the previousdistillation). The distillation took approximately 1 hour. The vaportemperature during the distillation was in the range of 54-64° C. andthe internal temperature was 67° C. at the end of the distillation. Themixture was slowly cooled to room temperature and stirred for anadditional 3 hours. The solids were collected by filtration. The wetcake was washed with 16.7% (v/v) of isopropyl alcohol in n-heptane(384.0 mL), followed by n-heptane (280.0 mL), and dried under vacuum at55° C. to provide 36.1 grams of the desired product as white solids in90% yield. The chemical purity is 99.79% by HPLC analysis. The chiralpurity is 99.8% by chiral HPLC analysis. ¹H NMR (499.7 MHz, DMSO-d6) δ(ppm): 12.21 (s, 1H), 10.71 (s, 3H), 8.80 (s, 1H), 8.72 (s, 1H), 8.40(s, 1H), 7.60 (d, J=3.5 Hz, 1H), 7.00 (d, J=3.5 Hz, 1H), 4.51 (td,J=9.75, 4.0 Hz, 1H), 3.25 (dd, J=17.3, 9.75 Hz, 1H), 3.14 (dd, J=17.0,4.0 Hz, 1H), 2.43-2.35 (m, 1H), 1.79-1.73 (m, 1H), 1.58-1.42 (m, 3H),1.41-1.33 (m, 1H), 1.30-1.23 (m, 2H), 1.19-1.12 (m, 1H); ¹³C NMR (125.7MHz, DMSO-d6) δ (ppm): 152.8, 151.2, 150.3, 140.0, 131.8, 127.7, 120.8,118.8, 113.5, 100.7, 63.3, 45.0, 29.8, 25.6, 25.0, 23.2; LCMS m/z:calculated for C₁₇H₁₈N₆ (M+H)⁺: =307.2. Found (M+H)⁺: 307.0.

4-(1H-Pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5)

Method B. To a reactor equipped with overhead stirring, condenser,thermowell, and nitrogen inlet was charged4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (4, 600 g,3.09 mol), toluene (4.2 L), and ethyl vinyl ether (334.5 g, 4.64 mol,0.44 L, 1.50 equiv) at room temperature before a solution of 2 M HCl indiethyl ether (39 mL, 0.078 mol, 0.025 equiv) was added dropwise. Theresulting reaction mixture was heated to 35-40° C. for 4-8 h. When HPLCanalysis showed that the reaction was deemed complete, the reactionmixture was cooled to 15-25° C. before being treated with an aqueousNaHCO₃ solution to pH>8. The two layers were separated, and the organiclayer was concentrated under reduced pressure to afford the crude1-(1-ethoxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(14), which was directly used in the subsequent Suzuki coupling reactionwithout further purification.

To a reactor equipped with overhead stirring, condenser, thermowell, andnitrogen inlet was charged water (H₂O, 1.5 L), potassium carbonate(K₂CO₃, 1047 g, 7.58 mol, 2.45 equiv),4-chloro-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(3a, 755 g, 2.66 mol), crude1-(1-ethoxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(14, 822 g based on 100% conversion, 3.09 mol, 1.16 equiv) made asdescribed above, and 1-propanol (6 L) at room temperature. The resultingreaction mixture was degassed three timed backfilling with nitrogen eachtime before being treated with tetrakis(triphenylphosphine)palladium(0)(9.2 g, 0.008 mol, 0.0026 equiv) at room temperature. The resultingreaction mixture was heated to gentle reflux (about 90° C.) for 1-4hours. When the reaction was deemed complete by HPLC, the reactionmixture was concentrated under reduced pressure to remove solvents. Theresidue was then cooled to room temperature, diluted with ethyl acetate(9 L) and water (4 L). The two layers were separated, and the aqueouslayer was extracted with ethyl acetate (2×2.5 L). The combined organiclayers were washed with water (2×2 L) and concentrated under reducedpressure to afford the crude4-(1-(1-ethoxyethyl)-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(15), which was directly used in the subsequent acid-promotedde-protection reaction without further purification.

To a reactor equipped with overhead stirring, condenser, thermowell, andnitrogen inlet was charged crude4-(1-(1-ethoxyethyl)-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(15, 1030.9 g based on 100% conversion, 2.66 mol), tetrahydrofuran (THF,0.9 L), water (H₂O, 4.4 L), and a 10% aqueous HCl solution (2.7 L, 10.64mol, 3.44 equiv) at room temperature. The resulting reaction mixture wasstirred at room temperature for 2-5 h. When the reaction was deemedcomplete by HPLC analysis, the reaction mixture was treated with a 30%aqueous sodium hydroxide (NaOH) solution (940 mL, 11.70 mol, 3.78 equiv)at room temperature. The resulting reaction mixture was stirred at roomtemperature for 1-2 h. The solids were collected by filtration, washedwith water (2×0.75 L), and dried in a vacuum oven at 45-55° C. toconstant weight to afford the crude4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 826.8 g, 839.1 g theoretical, 98.5% yield) as off-white solids (94.2area % pure by HPLC). This crude material was subsequentlyrecrystallized in acetonitrile to afford pure compound 5 (738.4 g, 839.1g theoretical, 88% yield) as white crystals (99.5 area % by HPLC), whichwas found to be identical in every comparable aspect to the materialmade from Method A.

4-(1H-Pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5)

Method C. To a reactor equipped with overhead stirring, condenser,thermowell, and nitrogen inlet was charged water (H₂O, 9.0 L), potassiumcarbonate (K₂CO₃, 4461 g, 32.28 mol, 2.42 equiv),4-chloro-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(3a, 3597 g, 12.67 mol),1-(1-ethoxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(14, 3550, 13.34 mol, 1.05 equiv), and 1-butanol (27 L) at roomtemperature. The resulting reaction mixture was degassed three timedbackfilling with nitrogen each time before being treated withtetrakis(triphenylphosphine)palladium(0) (46 g, 0.040 mol, 0.003 equiv)at room temperature. The resulting reaction mixture was heated to gentlereflux (about 90° C.) for 1-4 hours. When the reaction was deemedcomplete by HPLC, the reaction mixture was cooled to room temperaturebefore being filtered through a Celite bed. The Celite bed was washedwith ethyl acetate (2×2 L) before the filtrates and washing solutionwere combined. The two layers were separated, and the aqueous layer wasextracted with ethyl acetate (12 L). The combined organic layers wereconcentrated under reduced pressure to remove solvents, and the crude4-(1-(1-ethoxyethyl)-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(15) was directly charged back to the reactor with tetrahydrofuran (THF,4.2 L) for the subsequent acid-promoted de-protection reaction withoutfurther purification.

To a suspension of crude4-(1-(1-ethoxyethyl)-1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(15) made as described above in tetrahydrofuran (THF, 4.2 L) in thereactor was charged water (H₂O, 20.8 L), and a 10% aqueous HCl solution(16.2, 45.89 mol, 3.44 equiv) at room temperature. The resultingreaction mixture was stirred at 16-30° C. for 2-5 h. When the reactionwas deemed complete by HPLC analysis, the reaction mixture was treatedwith a 30% aqueous sodium hydroxide (NaOH) solution (4 L, 50.42 mol,3.78 equiv) at room temperature. The resulting reaction mixture wasstirred at room temperature for 1-2 h. The solids were collected byfiltration and washed with water (2×5 L). The wet cake was charged backto the reactor with acetonitrile (21.6 L), and resulting suspension washeated to gentle reflux for 1-2 h. The clear solution was then graduallycooled to room temperature with stirring, and solids were precipitatedout from the solution with cooling. The mixture was stirred at roomtemperature for an additional 1-2 h. The solids were collected byfiltration, washed with acetonitrile (2×3.5 L), and dried in oven underreduced pressure at 45-55° C. to constant weight to afford4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 3281.7 g, 3996.8 g theoretical, 82.1% yield) as white crystallinesolids (99.5 area % by HPLC), which was found to be identical in everycomparable aspect to the material made from Method A and B.

4-(1H-Pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5)

Method D. To a suspension of sodium hydride (NaH, 60 wt % oildisposition, 4.05 g, 101.3 mmol, 1.54 equiv) in 1,2-dimethoxyethane(DME, 20.0 mL, 192.4 mmol) at 0-5° C. (ice bath) was added4-chloropyrrolo[2,3-d]pyrimidine (1, 10.08 g, 65.6 mmol) in1,2-dimethoxyethane (DME, 80.0 mL, 769.6 mmol) slowly so that thetemperature was below 5° C. (−7° C. to 5° C.). A large amount of gas wasevolved immediately. The resulting reaction mixture was stirred at 0-5°C. for 30 min before trimethylsilylethoxymethyl chloride (2, 12.56 g,75.3 mmol, 1.15 equiv) was added slowly while the reaction temperaturewas maintained at <5° C. After the addition, the reaction was stirred at0° C. for 1 h before being warmed to room temperature for 23 h. When theHPLC and TLC showed that the reaction was deemed complete, the reactionmixture was quenched with water (46 mL) at room temperature, and thequenched reaction mixture, which contains the desired product (3a), wascarried into the next Suzuki coupling reaction directly without furtherwork-up and purification.

To the quenched reaction mixture, which contains crude4-chloro-7-[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidine(3a, 18.63 g, 65.64 mmol) from previous reaction as described above, wasadded 1,2-dimethoxyethane (DME, 38 mL), powder potassium carbonate(K₂CO₃, 23.56 g, 170.5 mmol, 2.6 equiv),1-(1-ethoxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(14, 18.60 g, 69.89 mmol, 1.06 equiv) at room temperature. The resultingmixture was degassed four times backfilling with nitrogen gas each timebefore being treated with tetrakis(triphenylphosphine)palladium(0)(244.2 mg, 0.21 mmol, 0.003 equiv) at room temperature. The resultingreaction mixture was degassed four times backfilling with nitrogen gaseach time before being warmed to 80° C. for 4-8 h. When TLC and HPLCshowed that the reaction was deemed complete, the reaction mixture wasgradually cooled to room temperature and filtered through a short bed ofCelite (10 g). The Celite bed was washed with ethyl acetate (EtOAc, 20mL). The two layers of the filtrate were separated, and the aqueouslayer was extracted with ethyl acetate (2×30 mL). The combined organicextracts were washed with saturated aqueous NaCl solution (20 mL), driedover magnesium sulfate (MgSO₄), and concentrated under reduced pressure.The residue, which contains the crude desired Suzuki coupling product(15), was then transferred to a 500 mL round bottom flask with THF (22mL) for subsequent de-protection reaction without further purification.

A solution of crude Suzuki coupling product (15) in THF (22 mL) wastreated with water (108 mL) and a solution of 10% aqueous HCl preparedby mixing 19.6 mL of concentrated HCl with 64 mL of H₂O at roomtemperature. The resulting reaction mixture was stirred at roomtemperature for 4-6 h. When TLC and HPLC showed the de-protectionreaction was deemed complete, a 30% aqueous sodium hydroxide (NaOH)solution prepared by dissolving 10.4 g of NaOH in 21.0 mL of H₂O wasadded slowly to the reaction mixture while maintaining the temperaturebelow 25° C. The solid gradually dissolved and re-precipitated after 10min. The mixture was stirred at room temperature for 1-2 h before thesolids were collected by filtration and washed with H₂O (50 mL). The wetcake was transferred to a 250 mL three-necked flask and treated withacetonitrile (MeCN, 112 mL) at room temperature. The mixture was heatedto reflux for 2 h before being cooled gradually to room temperature andstirred at room temperature for 1 h. The solids were collected byfiltration, washed with MeCN (36 mL) and dried at 40-45° C. in a vacuumoven to afford4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 15.3 g, 20.7 g theoretical, 73.9% yield) as white crystalline solids(99.4 area % by HPLC), which was found to be identical in everycomparable aspect to the material made from Method A, B, and C.

Racemic3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound)

Method B. Into a four-neck 250 mL round bottom flask equipped with astir bar, thermocouple, condenser and nitrogen inlet was charged(3S)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((S)-10, 13.9 g, 31.5 mmol), acetonitrile (84 mL) and3-cyclopentylacrylonitrile (8, a mixture of E and Z isomers, 3.82 g,31.5 mmol, 1.0 equiv) at room temperature. The resulting mixture wasthen treated with cesium carbonate (Cs₂CO₃, 2.57 g, 7.88 mmol, 0.25equiv) at room temperature. The reaction mixture was warmed to 65° C.and checked after 12 hours by chiral HPLC to determine the enantiomericratio of compound (R)-10 to compound (S)-10. When the ratio of compound(R)-10 to compound (S)-10 reached to one to one, the reaction mixturewas then allowed to cool to room temperature gradually and stirred atroom temperature for 24 to 48 h. The reaction mixture was monitored byHPLC to determine the level of4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5). The reaction was considered complete when the level of compound 5was found to be ≦2% by HPLC area %. The reaction mixture was thenfiltered through a Celite pad to remove insoluble solids present in thereaction solution. The filtrates were then concentrated under reducedpressure to remove about 40 mL of solvent. The concentrated solution wasdiluted with ethyl acetate (40 mL) and washed with 1 N aqueous HClsolution (40 mL). The two layers were separated, and the aqueous acidwash solution was back extracted with ethyl acetate (20 mL). Thecombined organic fractions were washed with 1 M aqueous sodiumbicarbonate (NaHCO₃) solution (45 mL) and 20% (w/w) brine solution (40mL). The organic fraction was dried over magnesium sulfate (MgSO₄) andconcentrated under reduced pressure to afford the crude racemic3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound, 13.6 g, 13.9 g theoretical, 97.8%)as an amber oil, which was found to be identical to the material made byMethod A. This crude product was found to be pure enough (>96 area % byHPLC) and was directly used in the subsequent chiral separation withoutfurther purification.

4-(1H-Pyrazol-4-yl)-7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5)

Method E. Into a 22 L four-neck flask equipped with overhead stirring,thermocouple, 2 L addition funnel and nitrogen inlet was charged(3S)-3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((S)-10, 491 g, 1.11 mol) and acetonitrile (4.5 L) at room temperature.The mixture was cooled to 0-10° C. before being treated dropwise with a1M solution of potassium tert-butoxide in THF (KO^(t)Bu, 2.0 L, 2.0 mol,1.8 equiv) via the addition funnel over 1.5 hours. Following theaddition of base the reaction mixture was allowed to return to roomtemperature and was stirred at room temperature for 12-24 h. When LC/MSshowed the reaction was deemed complete, the reaction mixture wasdiluted with ethyl acetate (EtOAc, 6 L) and 50% (w/w) aqueous ammoniumchloride solution (NH₄Cl, 4 L). The two layers were separated, and theaqueous fraction was back extracted with ethyl acetate (2 L). Thecombined organic fractions were washed with water (2 L) and brine (3 L),dried over magnesium sulfate (MgSO₄), and concentrated under reducedpressure to afford the crude4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 354 g, 350.1 g theoretical, 101.1% yield) as an amber oil, whichsolidified upon standing at room temperature in vacuo. This crudematerial was subsequently recrystallized in acetonitrile to afford purecompound 5 (308 g, 350.1 g theoretical, 88% yield) as white crystals(99.5 area % by HPLC), which was found to be identical in everycomparable aspect to the material made from Method A, B, C, and D.

(2S,3S)-2,3-Bis(benzoyloxy)succinicacid-(3R)-cyclopentyl-3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile(1:1; 17)

To a 250 ml round bottom flask equipped with a stir bar and nitrogeninlet was charged racemic3-cyclopentyl-3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile(9, 6.92 g, 0.0158 mol), acetonitrile (89.0 mL, 1.70 mol),tetrahydrofuran (15 mL, 0.185 mol) and acetone (15.0 mL, 0.204 mol) atroom temperature. The resulting solution was warmed to 50° C. beforebeing treated with (+)-2,3-dibenzoyl-D-tartaric acid (16, 8.52 g, 0.0238mol, 1.5 equiv) in one portion. The resulting homogeneous solution wasthen stirred at 50° C. for 10 minutes before being cooled gradually toroom temperature and stirred at room temperature for 21 hours. Thesolids were then collected by filtration, rinsed with a small volume ofhexane, and dried under reduced pressure to afford(2S,3S)-2,3-bis(benzoyloxy)succinicacid-(3R)-cyclopentyl-3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile(1:1; 17, 6.85 g, 12.6 g theoretical, 54% yield) as white crystals. Theenantiomeric purity of the isolated salt was analyzed by chiral HPLC andfound to be 74:26 favoring the desired R-enantiomer. For 17: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.86 (s, 1H), 8.78 (s, 1H), 8.42 (s, 1H), 8.04(dd, 4H, J=1.1, 8.4 Hz), 7.80 (d, 1H, J=3.5 Hz), 7.76 (tt, 2H, J=7.5,1.3 Hz), 7.73 (dd, 4H, J=7.9, 7.4 Hz), 7.12 (d, 1H, J=3.7 Hz), 5.90 (s,2H), 5.66 (s, 2H), 4.55 (td, 1H, J=4.2, 9.6 Hz), 3.54 (t, 2H, J=7.8 Hz),3.30 (dd, 1H, J=10.1, 17.6 Hz), 3.22 (dd, 1H, J=4.2, 16.9 Hz), 2.43 (m,1H), 1.82 (m, 1H), 1.70-1.14 (m, 7H), 0.85 (t, 2H, J=7.8 Hz), −0.083 (s,9H).

(3R)-Cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-10)

Method B. To a 250 mL round bottom flask was charged enantiomericallyenhanced (2S,3S)-2,3-bis(benzoyloxy)succinicacid-(3R)-cyclopentyl-3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile(1:1, 17, 6.85 g, 0.00862 mol), ethyl acetate (EtOAc, 70 mL, 0.717 mol)and water (20 mL, 1.11 mol) at room temperature, and the resultingsolution was cooled to 12° C. before being treated with 3 N aqueoussodium hydroxide solution (NaOH, 10.7 ml, 0.0321 mol, 3.72 equiv) toadjust pH to 8-9. The two layers were separated, and the aqueous layerwas extracted with ethyl acetate (30 mL). The combined organic fractionswere washed with 20% aqueous brine (20 mL), dried over magnesiumsulfate, filtered and concentrate under reduced pressure to affordenantiomerically enhanced(3R)-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile((R)-10, 3.31 g, 3.76 g theoretical g theoretical, 88%) as colorlessoil, which was analyzed by chiral HPLC and found to be 74:26 favoringthe desired R-enantiomer. For (R)-10: ¹H NMR (CD₃OD, 300 MHz) δ ppm 8.77(s, 1H), 8.68 (s, 1H), 8.43 (s, 1H), 7.66 (d, 1H, J=3.7 Hz), 7.06 (d,1H, J=3.7 Hz), 5.7 (s, 2H), 4.53 (td, 1H, J=4.5, 10.2 Hz), 3.62 (dd, 2H,J=8.0, 16.0 Hz), 3.26 (dd, 1H, J=9.7, 17.2 Hz), 3.17 (dd, 1H, J=4.0,17.0 Hz), 2.59 (m, 1H), 1.97 (m, 1H), 1.80-1.25 (m, 7H), 0.92 (t, 2H,J=8.4 Hz), −0.03 (s, 9H); C₂₃H₃₂N₆OSi (MW, 436.63), LCMS (EI) m/e 437(M⁺+H).

{4-[1-(1-Ethoxyethyl)-1H-pyrazol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methylpivalate (18)

To a 250 mL round bottom flask equipped with a stir bar, condenser and3-way valve was charged 4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (3f, 30 g, 0.112 mol), 1,4-dioxane (300 mL, 4.0 mol),1-(1-ethoxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(14, 35.8 g, 0.134 mol, 1.2 equiv), water (150 mL, 8.3 mol) andpotassium carbonate (K₂CO₃, 61.9 g, 0.448 mol, 4.0 equiv) at roomtemperature. The resulting mixture was degassed four times back fillingwith nitrogen each time before being chargedtetrakis(triphenylphosphine)palladium(0) (5.0 g, 0.00433 mol, 0.039equiv). The reaction mixture was then degassed four times back fillingwith nitrogen each time before being warmed to 85° C. The reactionmixture was stirred at 85° C. for 2-5 h. When the reaction was deemedcomplete, the reaction mixture was allowed to cool to room temperaturebefore being diluted with 20% aqueous brine (250 mL) and ethyl acetate(250 mL). The two layers were separated, and the aqueous layer wasextracted with ethyl acetate (250 mL). The combined organic fractionswas washed with water and brine, dried over magnesium sulfate (MgSO₄),and concentrated under reduced pressure. The residue was purified byflash column chromatography (SiO₂, 25% to 40% ethyl acetate/hexanegradient elution) to afford{4-[1-(1-ethoxyethyl)-1H-pyrazol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methylpivalate (18) as a orange oil, which was directly used in the subsequentreaction assuming the theoretical yield. For 18: C₁₉H₂₅N₅O₃ (MW,371.43), LCMS (EI) m/e 372 (M⁺+H).

[4-(1H-Pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate(19)

Method A. To a 1 L round bottom flask equipped with a stir bar andnitrogen inlet was charged{4-[1-(1-ethoxyethyl)-1H-pyrazol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methylpivalate (18, theoretical amount 41.6 g, 0.112 mol) made as describedabove and tetrahydrofuran (THF, 610 mL, 7.5 mol) at room temperature,and the resulting mixture was treated with an 2.0 N aqueous solution ofhydrochloric acid (140 mL, 0.28 mol, 2.5 equiv) at room temperature. Theresulting reaction mixture was subsequently stirred at room temperaturefor overnight. When the reaction was deemed complete, the reactionmixture was cooled to 0-5° C. before pH was adjusted to 9-10 with a 3 Maqueous sodium hydroxide (NaOH) solution (95 mL). The mixture was thenextracted with ethyl acetate (2×300 mL) and the combined organicextracts were washed with 20% aqueous brine solution (250 mL), driedover magnesium sulfate (MgSO₄), and concentrated under reduced pressureto afford the crude product as off-white to light yellow solids. Thecrude product was treated with methyl t-butylether (MTBE, 200 mL) andthe slurry was warm to reflux for 30 minutes before being cooled to roomtemperature. The solids were collected by filtration and washed withMTBE (2×40 mL), dried under reduced pressure to afford[4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate(19, 30.5 g, 33.52 g theoretical, 91% for two steps) as white tooff-white solids. For 19: ¹H NMR (DMSO-d₆, 300 MHz) δ ppm 13.40 (br s,1H), 8.75 (s, 1H), 8.66 (s, 1H), 8.32 (s, 1H), 7.68 (d, 1H, J=3.8 Hz),7.11 (d, 1H, J=3.8 Hz), 6.21 (s, 2H), 1.06 (s, 9H); C₁₅H₁₇N₅O₂ (MW,299.33), LCMS (EI) m/e 300 (M⁺+H).

Racemic(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20)

Method A. 3-Cyclopentylacrylonitrile (8, 14.6 g, 0.12 mol, 1.20 equiv)and DBU (18.2 mL, 0.12 mol, 1.2 equiv) was added to a suspension of4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate (19,30.0 g, 0.1 mol) in acetonitrile (45 mL) at room temperature. Theresulting reaction mixture was heated to 50-60° C. for 17 hours (a clearsolution developed midway through heating) then to room temperature for8 hours. When LCMS analysis showed the reaction was deemed complete, thereaction mixture was concentrated under reduced pressure and the residuewas dissolved in 2 L of ethyl acetate. The resulting solution was washedwith water (3×200 mL), dried over sodium sulfate (Na₂SO₄) andconcentrated under reduced pressure to give the crude product (20) as athick oil. The crude product was then purified by flash chromatography(SiO₂, 0-50% EtOAc/hexanes gradient elution) to afford racemic(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20, 13.0 g, 42.14 g theoretical, 30.8% yield) as a whitesolid. For 20: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.84 (s, 1H), 8.78 (s,1H), 8.39 (s, 1H), 7.74 (d, 1H, J=3.7 Hz,), 7.11 (d, 1H, J=3.8 Hz), 6.23(s, 2H), 4.53 (ddd, 1H, J=9.9, 9.6, 4.2 Hz), 3.26 (dd, 1H, J=17.4, 9.9Hz), 3.19 (dd, 1H, J=17.2, 4.3 Hz), 2.41 (m, 1H), 1.87-1.13 (m, 8H),1.07 (s, 9H); C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e 421.4 (M⁺+H).

Method B. To a stirred suspension of[4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate(19, 158 mg, 0.50 mmol) and 3-cyclopentylacrylonitrile (8, 122 mg, 1.0mmol, 2.0 equiv) in dimethyl sulfoxide (DMSO, 1.0 mL, 14 mmol) at roomtemperature was added powder potassium carbonate (K₂CO₃, 10.4 mg, 0.075mmol, 0.15 equiv). The reaction mixture was then stirred at roomtemperature for 5 h. The reaction mixture became a clear solution in 2h. When LCMS showed the reaction was deemed complete, the reaction wasquenched with water (H₂O, 5 mL) and extracted with ethyl acetate (EtOAc,3×15 mL). The combined organic extracts were washed with saturatedaqueous NaCl solution (10 mL), dried over magnesium sulfate (MgSO₄), andconcentrated under reduced pressure. The residue was then purified byflash chromatography (SiO₂, 0-50% EtOAc/hexanes gradient elution) toafford racemic(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20, 172.6 mg, 210 mg theoretical, 82% yield) as a white solid.For 20: ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.87 (s, 1H), 8.30 (s, 1H), 8.29(s, 1H), 7.47 (d, 1H, J=3.9 Hz), 6.75 (d, 1H, J=3.9 Hz), 6.24 (s, 2H),4.25 (m, 1H), 3.12 (dd, 1H, J=17.0, 8.7 Hz), 2.95 (dd, 1H, J=17.0, 3.9Hz), 2.58 (m, 1H), 1.95 (m, 1H), 1.72-1.52 (m, 5H), 1.25 (m, 2H), 1.14(s, 9H); C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e 421.4 (M⁺+H).

(R)-(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-21)

A solution of racemic(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20, 5.2 g, 12.36 mmol) in a mixture of ethanol and hexanes(1:9 by volume) was injected into preparative HPLC system equipped withthe chiral column (30×250 mm) packed with a silicagel based packingcoated with cellulose tris(3,5-dimethylphenyl)carbamate (available fromDaicel Chemical Industries, Ltd. (Daicel) as “Chiralcel® OD-H” (5 μm)).The chiral column was eluted with mobile phase made by a mixture ofethanol (EtOH) and hexanes in a 1 to 9 volume ratio at a flow rate of 32mL/min at room temperature. The column elution was monitored by UV atwavelength 220 nm. Under these conditions, baseline separation of thetwo enantiomers was achieved and the retention times were 16.4 minutes(Peak 1, the undesired (S)-enantiomer (S)-21) and 21.0 minutes (Peak 2,the desired (R)-enantiomer (R)-21), respectively. Each injection was 1.4mL of feed solution at a concentration of 50 mg/mL and each run cyclewas 14 minutes by using stack injections. Total 75 injections were takenfor this separation process. Fractions for Peak 1 (the undesired(S)-enantiomer, (S)-21) and Peak 2 (the desired (R)-enantiomer, (R)-21)were collected separately from each injection, and fractions collectedfor each peak were concentrated under reduced pressure. The residue fromeach evaporator was further dried under high vacuum to constant weightto afford(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-21, 2.36 g, 2.6 g theoretical, 90.8% yield) from Peak 2 asoff-white solids and(S)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((S)-21, 2.4 g, 2.6 g theoretical, 92.3% yield) from Peak 1 asoff-white solids.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of POM-(R)-21 and (S)-21 by using a Chiralcel® OD-H column(4.6×50 mm, 5 μm) purchased from Chiral Technologies, Inc. The twoenantiomers ((R)-21 and (S)-21) are separated with a resolution greaterthan 3.5 by using a mobile phase made of 10% ethanol and 90% hexanes atroom temperature with a flow rate of 1 mL/min. The UV detectionwavelength is 220 nm. The retention times are 14.1 minutes for (S)-21and 18.7 minutes for (R)-21, respectively.

The quality of each enantiomer separated by preparative chiral HPLCincluding chemical purity (HPLC area %) and chiral purity (chiral HPLCarea %) was analyzed and their structures are confirmed by NMRs andLC/MS. For (R)-21: achiral purity (99.2 area % by HPLC detected at 220nm); chiral purity (99.6 area % by chiral HPLC; 99.2% ee); ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.84 (s, 1H), 8.78 (s, 1H), 8.39 (s, 1H), 7.74(d, 1H, J=3.7 Hz,), 7.11 (d, 1H, J=3.8 Hz), 6.23 (s, 2H), 4.53 (ddd, 1H,J=9.9, 9.6, 4.2 Hz), 3.26 (dd, 1H, J=17.4, 9.9 Hz), 3.19 (dd, 1H,J=17.2, 4.3 Hz), 2.41 (m, 1H), 1.87-1.13 (m, 8H), 1.07 (s, 9H);C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e 421.4 (M⁺+H). For (S)-21: achiralpurity (99.3 area % by HPLC detected at 220 nm); chiral purity (99.8area % by chiral HPLC; 99.6% ee); ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.84(s, 1H), 8.78 (s, 1H), 8.39 (s, 1H), 7.74 (d, 1H, J=3.7 Hz,), 7.11 (d,1H, J=3.8 Hz), 6.23 (s, 2H), 4.53 (ddd, 1H, J=9.9, 9.6, 4.2 Hz), 3.26(dd, 1H, J=17.4, 9.9 Hz), 3.19 (dd, 1H, J=17.2, 4.3 Hz), 2.41 (m, 1H),1.87-1.13 (m, 8H), 1.07 (s, 9H); C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e421.4 (M⁺+H).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-12, free base)

Method B. To a stirred solution of(4-{1-[(1R)-2-cyano-1-cyclopentylethyl]-1H-pyrazol-4-yl}-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-21, 376 mg, 0.894 mmol) in methanol (4.0 mL, 99 mmol) atroom temperature was added 1.0 M solution of sodium hydroxide in water(NaOH, 179 μL, 0.179 mmol, 2.0 equiv). The reaction mixture was stirredat room temperature for overnight (15 h). When LCMS showed the reactionwas done cleanly, the reaction mixture was quenched with water (10 mL)and saturated aqueous NaCl solution (20 mL), and extracted with EtOAc(2×10 mL). The combined organic layers were washed with brine, driedover magnesium sulfate, filtered and concentrated under reduced pressureto afford(3R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-12, free base, 274 mg, 274 mg theoretical, 100% yield) as a paleyellow foam, which was found to be identical as the material made fromMethod A.

[4-(1H-Pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate(19)

Method B. To a oven dried 3 L 4-neck round bottom flask equipped with astirring bar, septa, thermocouple, 500 mL addition funnel and nitrogeninlet was charged sodium hydride (NaH, 60 wt % in mineral oil, 32.82 g,0.82 mol, 1.20 equiv) and anhydrous 1,2-dimethoxyethane (DME, 500 mL,4.8 mol) and the resulting mixture was cooled to 0-3° C. To a oven dried1 L round bottom flask was charged 4-chloro-7H-pyrrolo[2,3-d]pyrimidine(1, 105.0 g, 0.684 mol) and 1,2-dimethoxyethane (DME, 750 mL, 7.2 mol)and the resulting slurry was then portion wise added to the suspensionof sodium hydride in DME via large bore canula over 30 minutes at 5-12°C. The resulting reaction mixture was heterogeneous. Following theaddition, the cold bath was removed and the mixture was gradually warmedto room temperature and allowed to stir at room temperature for 1 hourbefore being cooled to 0-5° C. Chloromethyl pivalate (pivaloyloxymethylchloride, POM-Cl, 112 ml, 0.752 mol, 1.1 equiv) was added dropwise intothe reaction mixture over 30 minutes with stirring at 0-5° C. Theaddition of chloromethyl pivalate was mildly exothermic and the reactiontemperature went up to as high as 14° C. After addition of chloromethylpivalate, the cooling bath was removed and the reaction mixture wasallowed to return to room temperature and stirred at room temperaturefor 90 min. When the reaction was deemed complete after confirmed byHPLC, the reaction was carefully quenched with water (100 mL). And thisquenched reaction mixture, which contains crude POM-protectedchlorodeazapurine (3f), was used in the subsequent Suzuki couplingreaction without further work-up and purification.

To the quenched reaction mixture, which contains crude POM-protectedchlorodeazapurine (3f) made as described above was added4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (14, 200 g,0.75 mol, 1.10 equiv) and potassium carbonate (K₂CO₃, 189 g, 1.37 mol,2.0 equiv) at room temperature. The resulting mixture was degassed bypassing a stream of nitrogen through the solution for 15 minutes beforebeing treated with tetrakis(triphenylphosphine)-palladium(0) (7.9 g,0.68 mmol, 0.01 equiv) and the resulting reaction mixture was heated atreflux (about 82° C.) for 10 hours. When the reaction was deemedcomplete by TLC (1:1 hexanes/ethyl acetate) and LCMS, the reactionmixture was cooled to room temperature, diluted with ethyl acetate (2 L)and water (1 L). The two layers were separated, and the aqueous layerwas extracted with ethyl acetate (500 mL). The combined organic layerswere washed with water (2×1 L) and brine (1 L) before being concentratedunder reduced pressure to afford crude{4-[1-(1-ethoxyethyl)-1H-pyrazol-4-yl]-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methylpivalate (18) as a pale-yellow oil, which was directly used in thesubsequent de-protection reaction without further purification.

To a solution of crude 18 in THF (1 L, 12.3 mol) was treated with a 4 Naqueous HCl solution (500 mL) at room temperature. The resultingreaction mixture was subsequently stirred at room temperature for 5 h.When the reaction was deemed complete, the reaction mixture was cooledto 0-5° C. before pH was adjusted to 9-10 with a 1M aqueous sodiumhydroxide (NaOH) solution (2 L). The mixture was concentrated underreduced pressure to remove most of THF and the resulting suspension wasstirred at room temperature for 2 h. The solids were collected byfiltration, washed with water (3×500 mL), and dried under reducedpressure to afford[4-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl]methyl pivalate(19, 157.5 g, 204.43 g theoretical, 77% yield for three steps) as whiteto off-white solids, which was found to be sufficiently pure (>98 area %by HPLC) to do the subsequent reaction without further purification. For19: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 13.42 (br s, 1H), 8.76 (s, 1H), 8.67(s, 1H), 8.33 (s, 1H), 7.68 (d, 1H, J=3.8 Hz), 7.11 (d, 1H, J=3.8 Hz),6.21 (s, 2H), 1.06 (s, 9H); ¹³C NMR (DMSO-d₆, 100 MHz) δ ppm 177.74,152.31, 152.09, 151.91, 139.52, 130.39, 120.51, 113.93, 101.91, 67.26,38.98, 27.26; C₁₅H₁₇N₅O₂ (MW, 299.33), LCMS (EI) m/e 300 (M⁺+H).

(2S,3S)-2,3-Bis(benzoyloxy)succinicacid-(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (1:1; 22)

A solution of racemic(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20, 200 mg, 0.47 mmol) in a mixture of acetonitrile,tetrahydrofuran, and acetone (4 mL, 6:1:1) at room temperature waswarmed to 50° C. before being treated with (+)-2,3-dibenzoyl-D-tartaricacid (16, 84 mg, 0.235 mmol, 0.5 equiv) in one portion. The resultinghomogeneous solution was then stirred at 50° C. for 10 minutes beforebeing cooled gradually to room temperature and stirred at roomtemperature for 23 hours. The solids were then collected by filtration,rinsed with a small volume of hexane, and dried under reduced pressureto afford (2S,3S)-2,3-bis(benzoyloxy)succinicacid-(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (1:1; 22, 145 mg, 183 mg theoretical, 79.2% yield) as whitecrystals. The enantiomeric purity of the isolated salt was analyzed bychiral HPLC and found to be in a ratio of 87:13 favoring the desiredR-enantiomer. For 22: C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e 421.4(M⁺+H).

(R)-(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-21)

Method B. A solution of enantiomerically enhanced(2S,3S)-2,3-bis(benzoyloxy)succinicacid-(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (1:1; 22, 120 mg, 0.154 mmol) in ethyl acetate (10 mL) andwater (5.0 mL) at room temperature was cooled to 12° C. before beingtreated with 2 N aqueous potassium carbonate solution (K₂CO₃, 0.39 mL,0.77 mmol, 5.0 equiv) to adjust pH to 8-9. The two layers wereseparated, and the aqueous layer was extracted with ethyl acetate (30mL). The combined organic fractions were washed with 20% aqueous brine(20 mL), dried over magnesium sulfate, filtered and concentrate underreduced pressure to afford enantiomerically enhanced(R)-(4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate ((R)-21, 55.7 mg, 64.8 mg theoretical, 86% yield) as whitesolids, which was analyzed by chiral HPLC and found to be in a ratio of87:13 favoring the desired R-enantiomer. For ((R)-21: ¹H NMR (DMSO-d₆,400 MHz) δ ppm 8.84 (s, 1H), 8.78 (s, 1H), 8.39 (s, 1H), 7.74 (d, 1H,J=3.7 Hz,), 7.11 (d, 1H, J=3.8 Hz), 6.23 (s, 2H), 4.53 (ddd, 1H, J=9.9,9.6, 4.2 Hz), 3.26 (dd, 1H, J=17.4, 9.9 Hz), 3.19 (dd, 1H, J=17.2, 4.3Hz), 2.41 (m, 1H), 1.87-1.13 (m, 8H), 1.07 (s, 9H); C₂₃H₂₈N₆O₂ (MW,420.51), LCMS (EI) m/e 421.4 (M⁺+H).

Racemic3-Cyclopentyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)propanenitrile(23)

To a 500 mL round bottom flask equipped with a stir bar, condenser andnitrogen inlet was charged 3-cyclopentylacrylonitrile (8, a mixture of Eand Z isomers, 8.46 g, 0.067 mol, 1.3 equiv), acetonitrile (242 mL, 4.64mol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (4,10.0 g, 0.0515 mol), and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 16.2ml, 0.108 mol, 2.1 equiv) at room temperature. The resulting solutionwas then warmed to reflux, and the reaction mixture was stirred atreflux for 18 hours. When the reaction was deemed complete, the reactionmixture was allowed to cool to room temperature followed byconcentration under reduced pressure. The residue was purified directlyby flash column chromatography (SiO₂, 0% to 30% ethyl acetate/hexanegradient elution) to afford3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile(23, 13.1 g, 16.2 g theoretical, 81%) as off-white solids. This racemicmixture was directly used for subsequent chiral column separation without further purification. For 23: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.07(d, 1H, J=0.53 Hz), 7.65 (s, 1H), 4.42 (td, 1H, J=19.2, 4.5 Hz), 3.14(dd, 1H, J=9.39, 17.2 Hz), 3.08 (dd, 1H, J=4.58, 17.2 Hz), 2.31 (m, 1H),1.75 (m, 1H), 1.62-1.32 (m, 4H), 1.29-1.01 (m, 15H); C₁₇H₂₆BN₃O₂ (MW,315.22) LCMS (EI) m/e 316 [M⁺+H].

(R)-3-cyclopentyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)propanenitrile((R)-24) and(S)-3-cyclopentyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)propanenitrile((S)-24)

A solution of racemic3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile(23, 13.1 g, 41.56 mmol) in a mixture of ethanol and hexanes (8:2 byvolume) was injected into preparative HPLC system equipped with a chiralcolumn (20×250 mm) packed with amylose tri(3,5-dimethylphenyl)carbamateimmobilized on silicagel (Chiralpak® IA) from Chiral Technologies Inc.The chiral column was eluted with mobile phase made by a mixture ofethanol (EtOH) and hexanes in a 1 to 9 volume ratio at a flow rate of 18mL/min at room temperature. The column elution was monitored by UV atwavelength 220 nm. Under these conditions, a baseline separation of thetwo enantiomers was achieved and the retention times were 7.0 minutes(Peak 1, the undesired (S)-enantiomer (S)-24) and 8.3 minutes (Peak 2,the desired (R)-enantiomer (R)-24), respectively. Each injection was 0.8mL of feed solution at a concentration of 100 mg/mL and each run cyclewas 14 minutes by using stack injections. Total 164 injections weretaken for this separation process. Fractions for Peak 1 (the undesired(S)-enantiomer, (S)-24) and Peak 2 (the desired (R)-enantiomer, (R)-24)were collected separately from each injection, and fractions collectedfor each peak were concentrated under reduced pressure. The residue fromeach evaporator was further dried under high vacuum to constant weightto afford(R)-3-cyclopentyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)propanenitrile((R)-24, 6.19 g, 6.55 g theoretical, 94.5% yield) from Peak 2 asoff-white solids and(S)-3-cyclopentyl-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)propanenitrile((S)-24, 6.08 g, 6.55 g theoretical, 92.8% yield) from Peak 1 asoff-white solids.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of compound 24 ((R)-24 and (S)-24) by using a Chiralpak® IAcolumn (4.6×50 mm, 5 μm) purchased from Chiral Technologies, Inc. Twoenantiomers ((R)-24 and (S)-24) are separated with a resolution greaterthan 3.0 by using a mobile phase made from 15% ethanol and 85% hexanesat room temperature with a flow rate of 1 mL/min. The UV detectionwavelength is 220 nm. The retention times are 6.4 minutes for (S)-24 and7.6 minutes for (R)-24, respectively.

The quality of each enantiomer separated by preparative chiral HPLCincluding chemical purity (HPLC area %) and chiral purity (chiral HPLCarea %) was analyzed and their structures are confirmed by NMRs andLC/MS. For (R)-24: achiral purity (98.8 area % by HPLC detected at 220nm); chiral purity (99.8 area % by chiral HPLC; 99.6% ee); ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.07 (d, 1H, J=0.53 Hz), 7.65 (s, 1H), 4.42(td, 1H, J=19.2, 4.5 Hz), 3.14 (dd, 1H, J=9.39, 17.2 Hz), 3.08 (dd, 1H,J=4.58, 17.2 Hz), 2.31 (m, 1H), 1.75 (m, 1H), 1.62-1.32 (m, 4H),1.29-1.01 (m, 15H); C₁₇H₂₆BN₃O₂ (MW, 315.22) LCMS (EI) m/e 316 (M⁺+H).For (S)-24: achiral purity (98.6 area % by HPLC detected at 220 nm);chiral purity (99.6 area % by chiral HPLC; 99.2% ee); ¹H NMR (DMSO-d₆,400 MHz) δ ppm 8.07 (d, 1H, J=0.53 Hz), 7.65 (s, 1H), 4.42 (td, 1H,J=19.2, 4.5 Hz), 3.14 (dd, 1H, J=9.39, 17.2 Hz), 3.08 (dd, 1H, J=4.58,17.2 Hz), 2.31 (m, 1H), 1.75 (m, 1H), 1.62-1.32 (m, 4H), 1.29-1.01 (m,15H); C₁₇H₂₆BN₃O₂ (MW, 315.22) LCMS (EI) m/e 316 [M⁺+H].

Racemic3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound)

Method C. Into a 25 ml round bottom flask equipped with a stir bar,condenser, thermocouple and 3-way valve was charged3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile(23, 0.697 g, 2.21 mmol, 1.3 equiv),4-chloro-7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidine(3a, 0.506 g, 1.69 mmol), 1,4-dioxane (4.44 mL), water (4.44 mL), andsodium bicarbonate (NaHCO₃, 0.666 g, 7.93 mmol, 4.7 equiv) at roomtemperature. The resulting mixture was degassed four times backfillingwith nitrogen each time before tetrakis(triphenylphosphine)palladium(0)(91.6 mg, 0.079 mmol, 0.047 equiv) was added. The resulting reactionmixture was degassed four times backfilling with nitrogen each time. Thereaction was then warmed to 90° C. for 2-6 h. When TLC and HPLC showedthat the coupling reaction was deemed complete, the reaction mixture wasallowed to cool to room temperature followed by dilution with water (5mL) and ethyl acetate (10 mL). The two layers were separated, and theaqueous layer was back extracted with ethyl acetate (10 mL). Thecombined organic fractions were washed with water (10 mL) and saturatedaqueous NaCl solution (10 mL), dried over magnesium sulfate (MgSO₄), andconcentrated under reduced pressure to give the crude product (9) as anamber oil. The crude product was purified by flash column chromatography(SiO₂, 0% to 40% ethyl acetate/hexane gradient elution) to affordracemic3-cyclopentyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(9, racemic SEM-protected compound, 617 mg, 737.9 mg theoretical, 83.6%yield) as a yellow oil. For 9: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.83 (s,1H), 8.75 (s, 1H), 8.39 (s, 1H), 7.77 (d, 1H, J=3.7 Hz), 7.09 (d, 1H,J=3.7 Hz), 5.63 (s, 2H), 4.53 (td, 1H, J=19.4, 4.0 Hz), 3.51 (t, 2H,J=8.1 Hz), 3.23 (dq, 2H, J=9.3, 4.3 Hz), 2.41 (m, 1H), 1.79 (m, 1H),1.66-1.13 (m, 7H), 0.81 (t, 2H, J=8.2 Hz), 0.124 (s, 9H); C₂₃H₃₂N₆OSi(MW, 436.63), LCMS (EI) m/e 437 (M⁺+H) and m/e 459 (M⁺+Na).

Racemic(4-(1-(2-Cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20)

Method B. Into a 50 ml round bottom flask equipped with a stir bar,condenser and 3-way valve connected to nitrogen and vacuum was charged(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methyl pivalate (3f, 700 mg,2.61 mmol),3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile(23, 935 mg, 2.97 mmol, 1.13 equiv), 1,2-dimethoxyethane (DME, 10 mL, 96mmol), water (5 mL, 0.28 mol) and potassium carbonate (K₂CO₃, 1.82 g,7.84 mmol, 3.0 equiv) at room temperature. The resulting reactionmixture was degassed three times back filling with nitrogen each timebefore being charged tetrakis(triphenylphosphine)palladium(0) (30 mg,0.026 mmol, 0.010 equiv). The resulting reaction mixture was degassedfour times back filling with nitrogen each time and then warmed to 82°C. The reaction mixture was stirred at 82° C. for 6 hours. When thereaction was deemed complete, the reaction mixture was cooled to roomtemperature before being diluted with ethyl acetate (45 mL) and water(10 mL). The resulting mixture was stirred until the majority of solidshad gone into solution. The two layers were separated, and the aqueouslayer was extracted with ethyl acetate (1×25 mL). The combined organicfractions were washed with aqueous brine (2×25 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by flash chromatography (SiO₂, 0-50% ethyl acetate/hexanegradient elution) to afford racemic4-(1-(2-cyano-1-cyclopentylethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)methylpivalate (20, 0.97 g, 1.1 g theoretical, 88.6% yield) as colorless oil,which solidified upon standing at room temperature in vacuo. For 20: ¹HNMR (CDCl₃, 300 MHz) δ 8.85 (s, 1H), 8.29 (s, 1H), 8.27 (s, 1H), 7.45(d, 1H, J=3.8 Hz,), 6.73 (d, 1H, J=3.8 Hz), 6.22 (s, 2H), 4.23 (ddd, 1H,J=10.0, 8.6, 4.0 Hz), 3.10 (dd, 1H, J=17.0, 8.6 Hz), 2.92 (dd, 1H,J=17.0, 4.0 Hz), 2.56 (m, 1H), 2.00-1.25 (m, 8H), 1.12 (s, 9H);C₂₃H₂₈N₆O₂ (MW, 420.51), LCMS (EI) m/e 421 (M⁺+H).

(3R)-Cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-12, free base). Method C.

To a 25 mL round bottom flask equipped with a stir bar, condenser, andthree-way valve connected to nitrogen and vacuum was charged4-chloro-7H-pyrrolo[2,3-d]pyrimidine (1, 154 mg, 1.00 mmol),(3R)-3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile((R)-24, 445 mg, 1.41 mmol, 1.41 equiv), 1,4-dioxane (2.78 mL, 35.6mmol), water (1.39 mL, 77.2 mmol), and potassium carbonate (K₂CO₃, 693mg, 5.02 mmol, 5.0 equiv) at room temperature. The resulting mixture wasdegassed three times back filling with nitrogen each time before beingcharged tetrakis(triphenylphosphine)palladium(0) (207 mg, 0.180 mmol,0.18 equiv). The resulting reaction mixture was degassed four times backfilling with nitrogen each time and then warmed to 95° C. The reactionmixture was stirred at 95° C. for 17 hours. When the reaction was deemedcomplete, the reaction mixture was cooled to room temperature beforebeing diluted with ethyl acetate (20 mL) and 20% aqueous brine (11 mL).The mixture was stirred vigorously at room temperature until themajority of solids had gone into solution. The two layers wereseparated, and the aqueous layer was extracted with ethyl acetate (20mL). The combined organic extracts were washed with saturated brine (10mL), dried over MgSO₄, filtered, and concentrated under reducedpressure. The residue was then purified by flash chromatography (SiO₂,0-100% ethyl acetate/hexanes gradient elution) to afford(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-c]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile((R)-12, 197 mg, 306.4 mg theoretical, 64.3% yield) as colorless oil,which was solidified upon standing at room temperature. For (R)-12: ¹HNMR (DMSO-d₆, 400 MHz) δ ppm 12.1 (bs, 1H), 8.80 (d, 1H, J=0.42 Hz),8.67 (s, 1H), 8.37 (s, 1H), 7.59 (dd, 1H, J=2.34, 3.51 Hz), 6.98 (dd,1H, J=1.40, 3.44 Hz), 4.53 (td, 1H, J=19.5, 4.63 Hz), 3.26 (dd, 1H,J=9.77, 17.2 Hz), 3.18 (dd, 1H, J=4.32, 17.3 Hz), 2.40 (m, 1H), 1.79 (m,1H), 1.65 to 1.13 (m, 7H); C₁₇H₁₈N₆(MW, 306.37) LCMS (EI) m/e 307[M⁺+H].

(S)-3-Cyclopentyl-3-(4-(7-(diethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((S)-25)

Into a 100 ml round bottom flask equipped with a stir bar, condenser and3-way valve connected to nitrogen and vacuum was charged4-chloro-7-(diethoxymethyl)-7H-pyrrolo[2,3-c]pyrimidine (3b, 3.30 g,0.0129 mol),(3S)-3-cyclopentyl-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]propanenitrile((S)-24, 5.12 g, 0.0146 mol, 1.13 equiv), 1,4-dioxane (33.4 mL, 0.428mol), water (16.7 mL, 0.929 mol) and potassium carbonate (K₂CO₃, 8.03 g,0.0581 mol, 4.5 equiv) at room temperature. The resulting reactionmixture was degassed three times back filling with nitrogen each timebefore being charged tetrakis(triphenylphosphine)palladium(0) (1.49 g,0.00129 mol, 0.10 equiv). The mixture was degassed four times backfilling with nitrogen each time and then warmed to 95° C. The reactionmixture was stirred at 95° C. for 21 hours. When the reaction was deemedcomplete, the reaction mixture was cooled to room temperature beforebeing diluted with ethyl acetate (45 mL) and water (20 mL). Theresulting mixture was stirred until the majority of solids had gone intosolution. The two layers were separated, and the aqueous layer wasextracted with ethyl acetate (50 mL). The combined organic fractionswere washed with 20% aqueous brine (50 mL), dried over MgSO₄, filteredand concentrated under reduced pressure. The residue was purified byflash chromatography (SiO₂, 0% to 50% ethyl acetate/hexane gradientelution) afford(3S)-3-cyclopentyl-3-{4-[7-(diethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-1H-pyrazol-1-yl}propanenitrile((S)-25, 4.11 g, 5.27 g theoretical, 78% yield) as colorless oil, whichwas solidified upon standing at room temperature. For (S)-25: ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.84 (s, 1H), 8.74 (s, 1H), 8.38 (s, 1H), 7.71(d, 1H, J=3.8 Hz,), 7.12 (d, 1H, J=3.8 Hz), 6.76 (s, 1H), 4.53 (td, 1H,J=19.4, 4.3 Hz), 3.68 (m, 2H), 3.52 (m, 2H), 3.26 (dd, 1H, J=9.6, 17.3Hz), 3.19 (dd, 1H, J=4.3, 17.2 Hz), 2.41 (m, 1H), 1.80 (m, 1H),1.63-1.09 (m, 13H); C₂₂H₂₈N₆O₂ (MW, 408.50), LCMS (EI) m/e 409 (M⁺+H).

3-Cyclopropylacrylonitrile (27)

A solution of diethyl cyanomethylphosphonate (7, 779.5 g, 4.4 mol, 1.1equiv) in dry tetrahydrofuran (THF, 5.75 L) was stirred under nitrogenin an ice-water-methanol bath before a solution of 1 M potassiumtert-butoxide in THF (KO^(t)Bu, 4.2 L, 4.2 mol, 1.05 equiv) was added atsuch a rate as to keep the temperature below 0° C. After addition ofpotassium tert-butoxide solution was complete, the stirring wascontinued at 0-5° C. for 1 h. and a solution of cyclopentanecarbaldehyde(26, 280 g, 4.0 mol) in dry THF (290 ml) was added at such a rate as tomaintain the temperature below 0° C. The cold bath was removed, and thereaction mixture was gradually warmed to room temperature and stirred atroom temperature for overnight. When the reaction was deemed complete,the reaction mixture was partitioned between MTBE (14 L), water (10 L)and brine (6 L). The organic phase was washed with brine (6 L). Theaqueous phase was extracted with methyl tert-butyl ether (MTBE, 10 L)and washed with brine (6 L). The combined organic extracts wereconcentrated under reduced pressure and the residue was distilled toafford 3-cyclopropylacrylonitrile (27, 342.7 g, 372.5 g theoretical, 92%yield) as a colorless oil, which was found to be a mixture of E- andZ-isomer. For 27: ¹H NMR (DMSO-d₆, 400 MHz, for E-isomer) δ ppm 6.33(dd, 1H, J=16.3, 10.3 Hz), 5.69 (d, 1H, J=16.4 Hz), 1.66 (m, 1H), 1.02(m, 1H,), 0.93 (m, 1H), 0.69 (m, 2H) and (for Z-isomer) δ ppm 6.05 (t,1H, J=10.8 Hz), 5.45 (d, 1H, J=9.7 Hz), 1.82 (m, 1H), 1.02 (m, 1H), 0.93(m, 1H), 0.69 (m, 2H); ¹³C NMR (DMSO-d₆, 100 MHz, for E-isomer) δ ppm160.9, 118.4, 95.4, 15.4, 8.64 and (for Z-isomer) δ ppm 160.0, 117.3,95.2, 14.8, 8.4; C₆H₇N (MW, 93.13), GCMS (EI) m/e 92 (M⁺−H).

Racemic3-Cyclopropyl-3-{4-[7-(2-trimethylsilanylethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]pyrazol-1-yl}propionitrile(28, Racemic SEM-protected compound)

To a suspension of4-(1H-pyrazol-4-yl)-7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 1.115 Kg, 3.54 mol, 1.0 equiv) in acetonitrile (11 L) was added3-cyclopropylacrylonitrile (27, 428.7 g, 4.60 mol, 1.3 equiv) and1,8-diazobicyclo[5.4.0]undec-7-ene (DBU, 55 mL, 0.37 mol, 0.105 equiv).The resulting reaction mixture was heated to gentle reflux forapproximate 18 hours. When HPLC and TLC showed the reaction was deemedcomplete, the reaction mixture, which was a clear solution, was cooledto room temperature before being concentrated under reduced pressure togive the crude Michael addition product (28) as a dark red oil. Thecrude product was then diluted with dichloromethane, divided into threeportions and absorbed onto silica gel (3×2 Kg). The crude productabsorbed on silica gel was purified by column chromatography on three 2Kg silica gel columns (packed in 87.5:12.5 heptanes/EtOAc and elutedwith 87.5:12.5 to 25:75 heptanes/EtOAc). The fractions containing thepure desired product (28) were combined and concentrated under reducedpressure, transferred to afford racemic3-cyclopropyl-3-{4-[7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-pyrazol-1-yl}-propionitrile(28, racemic SEM-protected compound, 1.310 Kg, 1.446 Kg theoretical,90.6% yield) as a amber syrup, which was used for chiral columnseparation without further purification. For 28: C₂₁H₂₈N₅OSi (MW,408.57), LCMS (EI) m/e 409 (M⁺+H).

(3R)-3-Cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-29) and(3S)-3-Cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((S)-29)

A slurry of 1.5 Kg of 20-micron Chiralcel® OD chiral stationary phase(CSP) made by Daicel in 3.0 L of isopropanol (IPA) was packed into aPROCHROM Dynamic Axial Compression Column LC110-1 (11 cm ID×25 cm L;Column Void Vol.: approximate 1.5 L) under 150 bar of packing pressure.The packed column was then installed no a Novasep Hipersep HPLC unit.The column and HPLC system were flushed with methanol (17 L) and themobile phase made by a mixture of isopropanol and hexane (2:8 by volume,17 L). The feed solution was then prepared by dissolving3-cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile(28, racemic SEM-protected compound, 2500 g, 6.119 mol) in the mobilephase to a concentration of 80 g/L. The feed solution (120 mL perinjection) was then sequentially injected into the preparative HPLCchiral column for separation. The chiral column was eluted with themobile phase at a flow rate of 570 mL/min at room temperature. Thecolumn elution was monitored by UV at wavelength 330 nm. Under theseconditions, a baseline separation of the two enantiomers was achieved.The cycle time for each injection was 11 minutes and a total of 261injections were performed for this separation process. Fractions forPeak 1 (the undesired (S)-enantiomer, (S)-29) and Peak 2 (the desired(R)-enantiomer, (R)-29) were collected separately from each injection,and fractions collected for each peak were continuously concentrated at40° C. under reduced pressure (40-120 bar). The residue from eachevaporator was further dried under high vacuum to constant weight toafford(3R)-3-cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-29, 1150 g, 1250 g theoretical, 92%) from Peak 2 as a light yellowoil which solidified upon standing at room temperature in vacuo and(3S)-cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((S)-29, 1200 g, 1250 g theoretical, 96%) from Peak 1 as an yellow oilwhich solidified upon standing at room temperature in vacuo.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of SEM-protected compound ((R)-29 and (S)-29) using aChiralcel® OD-H column (4.6×250 mm, 5 μm), purchased from ChiralTechnologies, Inc. The two enantiomers of SEM-protected compound areseparated with a resolution greater than 4.0 by using a mobile phase of15% ethanol and 85% hexanes at room temperature with a flow rate of 1mL/min. The UV detection wavelength is 220 nm. The retention times for(S)-enantiomer ((S)-29) and (R)-enantiomer ((R)-29) are 9.4 minutes and12.4 minutes, respectively.

The quality of each enantiomer separated by preparative chiral HPLCincluding chemical purity (HPLC area %) and chiral purity (chiral HPLCarea %) was analyzed and their structures are confirmed by NMRs andLC/MS. For (R)-29: achiral purity (99.1 area % by HPLC detected at 220nm); chiral purity (99.4 area % by chiral HPLC; 98.8% ee); C₂₁H₂₈N₅OSi(MW, 408.57), LCMS (EI) m/e 409 (M⁺+H). For (S)-29: achiral purity (98.5area % by HPLC detected at 220 nm); chiral purity (99.2 area % by chiralHPLC; 98.4% ee); C₂₁H₂₈N₅OSi (MW, 408.57), LCMS (EI) m/e 409 (M⁺+H).

(3R)-3-Cyclopropyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-30)

A solution of(3R)-3-cyclopropyl-3-{4-[7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-pyrazol-1-yl}-propionitrile((R)-29, 102 g, 0.25 mol, 1.0 equiv) in MeCN (900 mL) and H₂O (75 mL)was treated with solid lithium tetrafluoroborate (LiBF₄, 186.0 g, 2.0mol, 8.0 equiv) in portions (the reaction temperature increased from 15to 38° on addition). The resulting reaction mixture was then heated atgentle reflux (light suspension formed) for 20 h. When LCMS showed thecleavage of the SEM group was complete, the reaction mixture was cooledto room temperature and subsequently to 12° before being adjusted to pH9-10 with addition of an aqueous NH₄OH solution (20%, 80 mL). Theresulting suspension was stirred at room temperature until LCMS showedno N-hydroxymethyl intermediate (M⁺+H=309) remained, typically within24-36 h. During this period the pH of the reaction mixture dropped to7-8, additional aqueous NH₄OH solution (20%) was added to readjust themixture to pH 9-10. The mixture was diluted with acetonitrile (300 mL),filtered, washing solids with acetonitrile (500 mL). The turbid filtratewas concentrated under reduced pressure to remove most of the MeCN togive a thick oil that contained some solids. The mixture was slowlydiluted with H₂O (500 mL) and the turbid solution was seeded. Thesolution was then concentrated under reduced pressure at roomtemperature until a thick suspension had formed. The suspension wasfurther diluted with H₂O (1 L) and the resulting suspension was stirredat room temperature for 2 h. The solids were collected by filtration,washed with H₂O (2×500 mL) and suction dried on funnel for 1.5 h. ¹⁹FNMR showed a small amount of inorganic fluoride present and TLC (5%MeOH/EtOAc) showed a small amount of baseline material existed.Therefore, the crude solids were re-slurried in H₂O (1 L) withmechanical stirring for 1 h before being collected by filtration andwashed with H₂O (500 mL). The wet cake was suction dried on the funnelfor 1.5 h then dried in a vacuum oven at 45-50° C. for 16 h to give(3R)-3-cyclopropyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-30, 60.8 g, 69.6 g theoretical, 87.4% yield) as a off-white solid.For (R)-30: C₁₅H₁₄N₆ (MW, 278.31), LCMS (EI) m/e 279 (M⁺+H).

(3R)-3-Cyclopropyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt ((R)-31, Phosphate)

A suspension of(3R)-3-cyclopropyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrile((R)-30, 60.0 g, 0.2158 mol, 1.0 equiv) in isopropanol (IPA, 900 mL) washeated to 77° C. to give a clear pale yellow solution. A solution ofcrystalline H₃PO₄ (23.3 g, 0.2374 mol, 1.1 equiv) in IPA (200 mL) wasadded in a steady stream from an addition funnel at 77-79° C., rinsingthe addition funnel with IPA (25 mL). An immediate turbidity wasdeveloped followed by formation of a white suspension. After about halfamount of the H₃PO₄ solution had been added the suspension becameextremely thick. An additional amount of IPA (100 mL) was added tofacilitate stirring. When addition was complete the suspension washeated at 75° C. for 1 h with the suspension becoming more mobile butremaining very thick. The suspension was cooled to room temperature over1 h and the solids were collected by filtration and washed with 50%IPA/heptane (750 mL) and dried. The solids were triturated with heptane(1.2 L) with stirring for overnight before being collected by filtrationand washed with heptane (300 mL) and dried in vacuum oven at 40-50° C.to constant weight to afford(3R)-3-cyclopropyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)pyrazol-1-yl]propionitrilephosphate salt ((R)-31, Phosphate, 76.7 g, 81.2 g theoretical, 94.5%yield) as a fine white crystalline solid. For (R)-31: ¹H NMR (DMSO-d₆,400 MHz) δ ppm 12.2 (bs, 1H), 9.62 (bs, 3H, H₃PO₄), 8.77 (s, 1H), 8.69(s, 1H), 8.39 (s, 1H), 7.59 (q, 1H, J=2.0 Hz), 6.98 (d, 1H, J=2.7 Hz),4.04 (m, 1H), 3.37 (dd, 1H, J=16.8, 8.0 Hz), 3.28 (dd, 1H, J=16.8, 5.1Hz), 1.43 (m, 1H), 0.68 (m, 1H), 0.49 (m, 3H); ¹³C NMR (DMSO-d₆, 100MHz) δ ppm 152.2, 150.9, 149.9, 139.3, 130.4, 127.0, 120.8, 118.1,112.9, 100.0, 62.6, 23.3, 15.7, 4.3, 3.8; C₁₅H₁₄N₆ (MW, 278.31), LCMS(EI) m/e 279.1 (M⁺+H).

Racemic4,4,4-Trifluoro-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile(33, Racemic SEM-protected compound)

To a flask equipped with a mechanical stirrer, nitrogen inlet andthermowell was added compound4-(1H-pyrazol-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine(5, 1424 g, 4.52 mol) and acetonitrile (14 L). The resulting suspensionwas added 4,4,4-trifluorocrotonitrile (32, 601.6 g, 4.97 mol, 1.1 equiv)followed by 1,8-diazobicyclo[5.4.0]undec-7-ene (DBU, 67 mL, 0.452 mol,0.1 equiv). A slight exotherm (5° C.) was noted upon the addition of theDBU. The reaction mixture was stirred at room temperature for 30 minuteswhen TLC and LCMS showed the reaction was deemed complete. The reactionmixture was then concentrated under reduced pressure to remove most ofthe solvent and the residue was purified by two silica gel columns (3 Kgeach) for chromatography purification. The column was eluting with 2:1heptane/ethyl acetate (30 L) followed by 1:1 heptane/ethyl acetate (30L). The fractions containing pure desired product (33) were combined andconcentrated under reduced pressure to afford racemic4,4,4-trifluoro-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile(33, Racemic SEM-protected compound, 1802 g, 1973 g theoretical, 91.3%yield) as a thick oil, which was directly used in subsequent chiralcolumn separation without further purification. For 33: ¹H NMR (DMSO-d₆,400 MHz) δ ppm 8.99 (s, 1H), 8.79 (s, 1H), 8.56 (s, 1H), 7.80 (d, 1H,J=3.7 Hz), 7.09 (d, 1H, J=3.7 Hz), 6.05 (m, 1H), 5.63 (s, 2H), 3.82 (dd,1H, J=17.5, 10.6 Hz), 3.66 (dd, 1H, J=17.0, 4.9 Hz), 3.50 (t, 2H, J=7.9Hz), 0.80 (t, 2H, J=8.2 Hz), −0.145 (s, 9H); ¹³C NMR (DMSO-d₆, 100 MHz)δ ppm 151.7, 151.3, 149.5, 140.8, 132.9, 130.4, 123.2 (J_(CF)=282 Hz),121.9, 116.2, 113.5, 100.2, 72.3, 65.7, 57.8 (J_(CF)=32.4 Hz), 17.1,−1.46; C₁₉H₂₃F₃N₆OSi (MW, 436.51), LCMS (EI) m/e 437 (M⁺+H).

(R)-4,4,4-Trifluoro-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile((R)-34) and(S)-4,4,4-Trifluoro-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile((S)-34)

A slurry of 1.5 Kg of 20-micron Chiralcel® OD chiral stationary phase(CSP) made by Daicel in 3.0 L of isopropanol (IPA) was packed into aPROCHROM Dynamic Axial Compression Column LC110-1 (11 cm ID×25 cm L;Column Void Vol.: approximate 1.5 L) under 150 bar of packing pressure.The packed column was then installed on a Novasep Hipersep HPLC unit.The column and HPLC system were flushed with methanol (17 L) and themobile phase made by a mixture of isopropanol and hexane (2:8 by volume,17 L). The feed solution was then prepared by dissolving4,4,4-trifluoro-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile(33, racemic SEM-protected compound, 3100 g, 7.1 mol) in the mobilephase to a concentration of 120 g/L. The feed solution (120 mL perinjection) was then sequentially injected into the preparative HPLCchiral column for separation. The chiral column was eluted with themobile phase at a flow rate of 570 mL/min at room temperature. Thecolumn elution was monitored by UV at wavelength 330 nm. Under theseconditions, a baseline separation of the two enantiomers was achieved.The cycle time for each injection was 11 minutes and a total of 210injections were performed for this separation process. Fractions forPeak 1 (the undesired (S)-enantiomer, (S)-34) and Peak 2 (the desired(R)-enantiomer, (R)-34) were collected separately from each injection,and fractions collected for each peak were continuously concentrated at40° C. under reduced pressure (40-120 bar). The residue from eachevaporator was further dried under high vacuum to constant weight toafford(3R)-3-cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((R)-34, 1457 g, 1550 g theoretical, 94%) from Peak 2 as a light yellowoil which solidified upon standing at room temperature in vacuo and(3S)-cyclopropyl-3-(4-(7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)propanenitrile((S)-34, 1488 g, 1550 g theoretical, 96%) from Peak 1 as an yellow oilwhich solidified upon standing at room temperature in vacuo.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of SEM-(R)-34 and (S)-34 using a Chiralcel® OD-H column(4.6×250 mm, 5 μm), purchased from Chiral Technologies, Inc. The twoenantiomers of SEM-protected compound are separated with a resolutiongreater than 9.0 by using a mobile phase of 15% ethanol and 85% hexanesat room temperature with a flow rate of 1 mL/min. The UV detectionwavelength is 220 nm. The retention times for (S)-enantiomer ((S)-34)and (R)-enantiomer ((R)-34) are 11.2 minutes and 22.2 minutes,respectively.

The quality of each enantiomer separated by preparative chiral HPLCincluding chemical purity (HPLC area %) and chiral purity (chiral HPLCarea %) was analyzed and their structures are confirmed by NMRs andLC/MS. For (R)-34: achiral purity (99.2 area % by HPLC detected at 220nm); chiral purity (99.4 area % by chiral HPLC; 98.8% ee); ¹H NMR(DMSO-d₆, 400 MHz) δ ppm 8.99 (s, 1H), 8.79 (s, 1H), 8.56 (s, 1H), 7.80(d, 1H, J=3.7 Hz), 7.09 (d, 1H, J=3.7 Hz), 6.05 (m, 1H), 5.63 (s, 2H),3.82 (dd, 1H, J=17.5, 10.6 Hz), 3.66 (dd, 1H, J=17.0, 4.9 Hz), 3.50 (t,2H, J=7.9 Hz), 0.80 (t, 2H, J=8.2 Hz), −0.145 (s, 9H); ¹³C NMR (DMSO-d₆,100 MHz) δ ppm 151.7, 151.3, 149.5, 140.8, 132.9, 130.4, 123.2 (J_(CF),=282 Hz), 121.9, 116.2, 113.5, 100.2, 72.3, 65.7, 57.8 (J_(CF)=32.4 Hz),17.1, −1.46; C₁₉H₂₃F₃N₆OSi (MW, 436.51), LCMS (EI) m/e 437 (M⁺+H). For(S)-34: achiral purity (99.1 area % by HPLC detected at 220 nm); chiralpurity (99.2 area % by chiral HPLC; 98.4% ee); ¹H NMR (DMSO-d₆, 400 MHz)δ ppm 8.99 (s, 1H), 8.79 (s, 1H), 8.56 (s, 1H), 7.80 (d, 1H, J=3.7 Hz),7.09 (d, 1H, J=3.7 Hz), 6.05 (m, 1H), 5.63 (s, 2H), 3.82 (dd, 1H,J=17.5, 10.6 Hz), 3.66 (dd, 1H, J=17.0, 4.9 Hz), 3.50 (t, 2H, J=7.9 Hz),0.80 (t, 2H, J=8.2 Hz), −0.145 (s, 9H); ¹³C NMR (DMSO-d₆, 100 MHz) δ ppm151.7, 151.3, 149.5, 140.8, 132.9, 130.4, 123.2 (J_(CF)=282 Hz), 121.9,116.2, 113.5, 100.2, 72.3, 65.7, 57.8 (J_(CF)=32.4 Hz), 17.1, −1.46;C₁₉H₂₃F₃N₆OSi (MW, 436.51), LCMS (EI) m/e 437 (M⁺+H).

4,4,4-Trifluoro-3(R)-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pyrazol-1-yl]-butyronitrile((R)-35)

To a flask equipped with a thermowell, reflux condenser, mechanicalstirrer, and nitrogen inlet was added4,4,4-trifluoro-3(R)-{4-[7-(2-trimethylsilanyl-ethoxymethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-pyrazol-1-yl}-butyronitrile((R)-34, 312 g, 0.716 mol), acetonitrile (4.5 L) and water (376 mL). Theresulting mixture was then treated with solid lithium tetrafluoroborate(LiBF₄, 697 g, 7.16 mol, 10.0 equiv) in portions at room temperature.The mixture was heated at reflux for 13 hours. When TLC indicated thatno starting material remained and two products (fully deprotected andthe hydroxymethyl analog) were produced, the reaction mixture was cooledto room temperature and then to 0° C. in an ice/water bath before beingtreated dropwise with an aqueous ammonium hydroxide solution (NH₄OH,20%, 245 mL) at 0-5° C. to bring the pH to between 9 and 9.5 asdetermined by 5-10 range pH strips. The ice bath was removed and thethick suspension was stirred at room temperature for overnight. WhenHPLC showed the reaction was complete, the reaction mixture was treatedwith water (1 L), brine (500 mL) and ethyl acetate (7 L). The two layerswere separated and the aqueous layer was extracted with ethyl acetate(2×2 L). The combined organic layers were concentrated under reducedpressure and the residue was re-dissolved in ethyl acetate (4 L) andwashed with brine (2×2 L). The organic layer was dried over sodiumsulfate, and the solvents were removed under reduced pressure to afforda thick slurry. Heptane was added to the thick slurry and solventremoval was continued until most of the ethyl acetate was removed. Thesolids were collected by filtration and dried in vacuum to afford thecrude product ((R)-35, 206 g, 219.3 g theoretical, 94% yield, 98% pureby HPLC) as white powders. The crude product was re-crystallized fromethanol (700 mL) to afford pure4,4,4-trifluoro-3(R)-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pyrazol-1-yl]-butyronitrile((R)-35, 188.6 g, 219.3 g theoretical, 86% yield, >99.5% pure by HPLC)as fine white crystalline solids. For (R)-35: ¹H NMR (DMSO-d₆, 500 MHz)δ ppm 12.2 (bs, 1H), 8.95 (s, 1H), 8.74 (s, 1H), 8.53 (s, 1H), 7.63 (d,1H, J=3.7 Hz), 6.97 (d, 1H, J=3.8 Hz), 6.04 (m, 1H), 3.81 (dd, 1H,J=17.1, 10.1 Hz), 3.65 (dd, 1H, J=17.1, 5.0 Hz); ¹³C NMR (DMSO-d₆, 125MHz) δ ppm 152.3, 151.0, 149.0, 140.7, 132.7, 127.2, 123.1 (J_(CF)=284Hz), 122.2, 116.2, 113.1, 99.5, 57.7 (J_(CF)=33.0 Hz), 17.3; C₁₃H₉F₃N₆(MW, 306.25), LCMS (EI) m/e 307 (M⁺+H).

3-[4-(7-{[2-(Trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile(37)

Into a 250 mL three-neck round bottom flask equipped with a stir bar,condenser, thermocouple and nitrogen inlet was charged4-(1H-pyrazol-4-yl)-7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidine(5, 10.3 g, 0.033 mol), 2-butenenitrile (36, 3.0 mL, 0.037 mmol, 1.12equiv) and acetonitrile (100 mL, 2.0 mol) at room temperature. Theresulting mixture was treated with 1,8-diazabicyclo[5.4.0]undec-7-ene(DBU, 2.0 mL, 0.013 mol, 0.4 equiv) and was subsequently warmed to 55°C. The reaction mixture was stirred at 55° C. for 15-20 h. When LC/MSshowed the reaction was deemed complete, the reaction mixture wasconcentrated under reduced pressure to yield an orange oil. The crudeproduct was then purified by flash column chromatography (SiO₂, 40-80%ethyl acetate/hexane gradient elution) to afford3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile(37, 12.3 g, 12.62 g theoretical, 97.5% yield) as a colorless oil, whichsolidified upon standing at room temperature in vacuo. For 37: ¹H NMR(CDCl₃, 400 MHz) δ ppm 8.84 (s, 1H), 8.33 (s, 1H), 8.30 (s, 1H), 7.39(d, 1H, J=3.8 Hz), 6.79 (d, 1H, J=3.8 Hz), 5.67 (s, 2H), 4.77 (m, 1H),3.53 (t, 2H, J=8.2 Hz), 3.05 (dd, 1H, J=16.8, 6.2 Hz), 2.98 (dd, 1H,J=16.8, 6.3 Hz), 1.79 (d, 3H, J=6.5 Hz), 0.91 (t, 2H, J=8.3 Hz), −0.068(s, 9H); C₁₉H₂₆N₆OSi (MW, 382.53), LCMS (EI) m/e 383 (M⁺+H).

(S)-3-(4-(7-((2-(Trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile((S)-38) and(R)-3-(4-(7-((2-(Trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile((R)-38)

A solution of racemic3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile(37, 38.3 g, 0.1 mmol) in a mixture of ethanol and hexanes (15:85 byvolume) was injected into preparative HPLC system equipped with achiralcolumn (30×250 mm) packed with silica gel coated with cellulosetri(3,5-dimethylphenyl carbamate) (Available at Chiral technologies Inc.as Chiralcel® OD-H, 5 μm). The column was eluted with mobile phase madeby a mixture of ethanol (EtOH) and hexanes in a 15 to 85 volume ratio ata flow rate of 32 mL/min at room temperature. The column elution wasmonitored by UV detection at a wavelength of 220 nm. Under theseconditions, a baseline separation of the two enantiomers was achievedand the retention times were 15.1 minutes (Peak 1, the undesired(R)-enantiomer (R)-38) and 19.6 minutes (Peak 2, the desired(S)-enantiomer (S)-38), respectively. Each injection was 0.5 mL of feedsolution at a concentration of 200 mg/mL and the cycle time for eachinjection was 14 minutes by using stack injections. A total of 384injections were performed for this separation process. Fractions forPeak 1 (the undesired (R)-enantiomer, (S)-38) and Peak 2 (the desired(S)-enantiomer, (S)-38) were collected separately from each injection,and fractions collected for each peak were concentrated under reducedpressure. The residue from each evaporator was further dried under highvacuum to constant weight to afford((S)-3-(4-(7-((2-trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile((S)-38, 17.43 g, 19.15 g theoretical, 91% yield) from Peak 2 asoff-white solids and(R)-3-(4-(7-((2-trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)butanenitrile((R)-38, 17.8 g, 19.15 g theoretical, 93% yield) from Peak 1 asoff-white solids.

A chiral HPLC method was developed for chiral purity evaluation of bothenantiomers of SEM-(R)-38 and (S)-38 by using a Chiralcel® OD-H column(4.6×250 mm, 5 μm), purchased from Chiral Technologies, Inc., packedwith a silicagel coated with cellulose tris(3,5-dimethylphenyl carbamate(Chiralcel® OD). The two enantiomers ((R)-38 and (S)-38) are separatedwith a resolution greater than 3.0 by using a mobile phase made from 15%ethanol and 85% hexanes at room temperature with a flow rate of 0.8mL/min. The UV detection wavelength is 220 nm. The retention times are17.8 minutes for (R)-38 and 21.5 minutes for (S)-38, respectively.

The quality of each enantiomer separated by preparative chiral HPLCincluding chemical purity (HPLC area %) and chiral purity (chiral HPLCarea %) was analyzed and their structures are confirmed by NMRs andLC/MS. For (S)-38: achiral purity (99.3 area % by HPLC detected at 220nm); chiral purity (99.5 area % by chiral HPLC; 99.0% ee); ¹H NMR(CDCl₃, 400 MHz) δ ppm 8.84 (s, 1H), 8.33 (s, 1H), 8.30 (s, 1H), 7.39(d, 1H, J=3.8 Hz), 6.79 (d, 1H, J=3.8 Hz), 5.67 (s, 2H), 4.77 (m, 1H),3.53 (t, 2H, J=8.2 Hz), 3.05 (dd, 1H, J=16.8, 6.2 Hz), 2.98 (dd, 1H,J=16.8, 6.3 Hz), 1.79 (d, 3H, J=6.5 Hz), 0.91 (t, 2H, J=8.3 Hz), −0.068(s, 9H); C₁₉H₂₆N₆OSi (MW, 382.53), LCMS (EI) m/e 383 (M⁺+H). For (R)-38:achiral purity (99.1 area % by HPLC detected at 220 nm); chiral purity(99.4 area % by chiral HPLC; 98.8% ee); ¹H NMR (CDCl₃, 400 MHz) δ ppm8.84 (s, 1H), 8.33 (s, 1H), 8.30 (s, 1H), 7.39 (d, 1H, J=3.8 Hz), 6.79(d, 1H, J=3.8 Hz), 5.67 (s, 2H), 4.77 (m, 1H), 3.53 (t, 2H, J=8.2 Hz),3.05 (dd, 1H, J=16.8, 6.2 Hz), 2.98 (dd, 1H, J=16.8, 6.3 Hz), 1.79 (d,3H, J=6.5 Hz), 0.91 (t, 2H, J=8.3 Hz), −0.068 (s, 9H); C₁₉H₂₆N₆OSi (MW,382.53), LCMS (EI) m/e 383 (M⁺+H).

(3S)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile((S)-39)

Into a 5 liter four neck round bottom flask equipped with overheadstirring, condenser, thermocouple and nitrogen inlet was charged(3S)-3-[4-(7-{[2-(trimethylsilyl)ethoxy]methyl}-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile((S)-38, 82.3 g, 0.215 mol), acetonitrile (1510 mL), water (135 mL) andsolid lithium tetrafluoroborate (LiBF₄, 206 g, 2.15 mol, 10.0 equiv).The resulting reaction mixture was warmed to reflux and stirred atreflux for 24-36 h. When HPLC and TLC showed that the reaction wasdeemed complete, the reaction mixture was cooled to room temperature. Anaqueous ammonium hydroxide (NH₄OH) solution (20% v/v) was added to thereaction mixture to adjust pH to 9-10. The resulting reaction mixturewas stirred at room temperature for 15-24 h. When HPLC and TLC showedthe de-protection reaction was deemed complete, the reaction mixture wasfiltered through a Celite pad to remove the insoluble materials. TheCelite pad was washed with ethyl acetate (500 mL). The filtrate wasfurther diluted with ethyl acetate (1 L) before being washed with a 20%sodium chloride (NaCl) aqueous solution (1 L). The aqueous fraction wasback extracted with ethyl acetate (2×500 mL). The combined organicfractions were then concentrated under reduced pressure to remove thesolvents to generate a thick white slurry. The slurry was treated withwater (2 L) and the resulting mixture was stirred at room temperaturefor 18 hours. The solids were collected by filtration, and the wet cakewas washed with methyl tert-butylether (MTBE, 500 mL) and heptane (500mL) before being dried at 50° C. in a vacuum oven to constant weight.The dried, crude product (45 g) was then re-crystallized in ethanol (500mL) and heptane (350 mL) to afford(3S)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]butanenitrile((S)-39, 42.8 g, 54.2 g theoretical, 79% yield) as white solids. For(S)-39: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 12.1 (bs, 1H), 8.76 (s, 1H),8.67 (s, 1H), 8.36 (s, 1H), 7.59 (d, 1H, J=3.5 Hz), 6.98 (d, 1H, J=3.5Hz), 4.98 (m, 1H), 3.19 (d, 2H, J=6.6 Hz), 1.57 (d, 3H, J=6.6 Hz);C₁₃H₁₂N₆ (MW, 252.27), LCMS (EI) m/e 253 (M⁺+H).

2-Cyano-4,4-diethoxy-butyric acid ethyl ester (4)

Bromoacetaldehyde diethylacetal (3, 541 g, 2.75 mol) was added to asuspension of powdered potassium carbonate (379.6 g, 2.75 mol, 1.0equiv) and sodium iodide (33 g, 0.22 mol, 0.08 equiv) in ethylcyanoacetate (2, 1.55 Kg, 13.75 mol, 5.0 equiv). Upon addition of thealdehyde to the reaction mixture, the resulting solution turned yellow.The reaction mixture was slowly heated to 140-150° C. collecting thevolatile material in a Dean Stark trap. This material was discarded.Fairly vigorous gas evolution was observed to begin at 140° C. Thereaction was monitored by G.C. and was observed to be near completion at90 minutes. Heating was continued for an additional 45 minutes when gasevolution was observed to have ceased. The reaction mixture was thencooled to room temperature and partitioned between 4 L water and 2 Lmethyl tert-butyl ether (MTBE). The layers were separated and theaqueous layer was extracted with an additional 2 L of MTBE. The aqueouslayer was checked for product by G.C. then discarded. The organic layerswere dried over sodium sulfate, filtered and concentrated in vacuum. Thecrude product was purified by fractional distillation (91-105° C. @0.53-0.65 mm/Hg) to afford 2-cyano-4,4-diethoxy-butyric acid ethyl ester(4, 359.4 g, 630.5 g theoretical, 57%) as a oil. For 4: ¹H NMR (DMSO-d₆,300 MHz) δ ppm 4.60 (t, 1H, J=5.6 Hz), 4.15 (m, 3H), 3.59 (m, 2H), 3.45(m, 1H), 2.11 (t, 2H, J=6.2 Hz), 1.22 (t, 3H, J=6.9 Hz), 1.10 (dt, 6H,J=7.1, 6.9 Hz).

7H-Pyrrolo[2,3-d]pyrimidin-4-ol (7)

Formamidine acetate (5, 1.04 Kg, 10 mol, 1.25 equiv) was added to 7.52 Lof (21% wt) sodium ethoxide (EtONa) in ethanol (EtOH, 62.5 equiv) andthe resulting solution was stirred for 60 minutes.2-cyano-4,4-diethoxy-butyric acid ethyl ester (4, 1.8 Kg, 8.0 mol) wasthen added and the resulting reaction mixture was refluxed for sevenhours. The stirring was turned off after the solution was cooled and thesolids were allowed to settle. The supernatant ethanol solution wasremoved, leaving the solids in the bottom of the reaction flask. Theethanol was evaporated and the residue was added back to the solidsremaining in the reaction flask with water and ice at a ratio of 600mL/mol. A solution of 6 N aqueous HCl was added to the resultingsolution at a ratio of 500 mL/mol at 15° C. The resulting solution wasthen heated at 45° C. for 45 minutes. The solution was again cooled to15° C. and the pH was adjusted to 8.0 with the addition of aqueousammonium hydroxide. The precipitated solids were collected byfiltration, washed with water (2×225 mL/mol) and pulled dry. The solidswere further washed with 1:1 ethyl acetate/heptane (500 mL/mol), thenheptane (2×250 mL/mol) and dried in vacuum to afford7H-pyrrolo[2,3-d]pyrimidin-4-ol (7, 738.6 g, 1081 g theoretical, 68.3%)as yellow to brown to yellow crystalline material. For 7: ¹H NMR(DMSO-d₆, 300 MHz) δ ppm 11.88 (bs, 1H), 11.80 (bs, 1H), 7.81 (s, 1H),7.02 (dd, 1H, J=3.2, 2.3 Hz), 6.42 (dd, 1H, J=3.5, 2.3 Hz); C₆H₅N₃O (MW,135.12), LCMS (EI) m/e 136 (M⁺+H) and (M⁺+Na) m/e 158.

4-Chloro-7H-pyrrolo[2,3-d]pyrimidine (1)

4-Hydroxy-7H-pyrrolo[2,3-d]pyrimidine (7, 306 g, 2.25 mol) was added inportions over 20 min to phosphorus oxychloride (1050 ml, 1727 g, 11.26mol, 5.0 equiv). Stirring was continued at room temperature for 15 minthen this suspension was slowly heated to reflux and the evolvinghydrochloric acid was scrubbed through 20% sodium hydroxide solution.Reflux was continued for 30 min after all material went in solution. Thereaction mixture was allowed to cool to 60° C. and it was poured ontoice (5 Kg) with stirring. Stirring was continued for 20 min andpotassium carbonate was slowly added in portions to adjust pH to 7.5.Ice was added as needed to keep the temperature below 20° C. Theprecipitate was collected by filtration, washed well with water anddried in a vacuum oven (30° C.). The crude material was taken in ethylacetate and stirred at 50° C. for 1.5 hrs. This solution was treatedwith charcoal, stirred at 50° C. for an additional 20 min and filteredhot through celite. The resulting solution was concentrated to 900 mland cooled in an ice bath with stirring. The precipitate was collectedby filtration, washed with small volume of cold ethyl acetate and driedin a vacuum oven (40° C.) to afford 4-chloro-7H-pyrrolo[2,3-d]pyrimidine(1, 227 g, 334.8 g theoretical, 67.8%) as yellow to brown crystallinesolids. Further concentration of the mother liquor produces anadditional crop of the desired product (5-10%) as yellow to browncrystals of less purity. For 1: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 12.58(bs, 1H), 8.58 (s, 1H), 7.69 (d, 1H, J=3.5 Hz), 6.59 (d, 1H, J=3.5 Hz);C₆H₄ClN₃ (MW, 153.57), LCMS (EI) m/e 154/156 (M⁺+H).

4,6-Dichloropyrimidine-5-carbaldehyde (9)

In a 5 L 4-neck flask equipped with mechanical stirrer, addition funnel,condenser, thermocouple, and a N₂ sweep into an aqueous NaOH scrubbingsolution, phosphorous oxychloride (1 L, 10.572 mol, 4.82 equiv) wascooled in an ice/salt bath. N,N-Dimethylformamide (DMF, 320 mL, 4.138mol, 1.85 equiv) was added dropwise at 0±2° C. After addition of ˜100 mLof DMF (˜0.5 hr) crystallization occurred and the reaction temperaturewas increased from 0 to 10° C. Addition was stopped and the mixture wasallowed to recool to ˜2° C. The remaining DMF was added over 2.5 hr at<8° C. The suspension became very thick making stirring difficult. Whenaddition of DMF was complete, the mixture was stirred 0.5 hr at 3-5° C.4,6-dihydroxypyrimidine (8, 250 g, 2.232 mol) was added portion wise asa solid. After about one third of 4,6-dihydroxypyrimidine was added thereaction mixture became more mobile and a slow exothermic phenomenaoccurred with the reaction temperature increasing to ˜12° C. over 0.5hr. The remaining 4,6-dihydroxypyrimidine was added portion wise over0.25 hr with the reaction temperature increasing from 12 to 27° C. Thereaction temperature was maintained at 25-27° C. with intermittentcooling during which time the yellow suspension became thinner, thenthicker once again. After the exothermic phenomenon subsided in about 1hr, the reaction mixture was heated slowly. At about 55° C. the reactionmixture became extremely thick and the second mild exothermic phenomenonwas occurred. The heating mantle was removed while the reactiontemperature continued to increase to about 63° C. and remained at thistemperature for several minutes before dropping. Heating of the mixturewas resumed until gentle reflux (about 100° C.) was attained. At about95° C. a steady, fairly rapid evolution of HCl began and the reactionmixture gradually thinned and darkened. After about 0.5 hr a clear,brown solution developed with the reflux temperature slowly increasingto 115° C. over 1.25 hr. After a total of 2.5 hr at reflux, the reactionmixture was cooled to room temperature and stirred overnight. ExcessPOCl₃ (as much as possible) was removed under reduced pressure (bathtemperature 45-50° C.). The thick residual brown oil was poured veryslowly into cold H₂O (5 L) in a 20 L separation funnel, adding ice asneeded to maintain the aqueous mixture near room temperature. Theaqueous mixture was extracted with EtOAc (2×3 L, 1×2 L). The combinedEtOAc extracts were washed with H₂O (2×2.5 L), saturated NaHCO₃ aqueoussolution (1 L), brine (1 L), dried over Na₂SO₄, filtered, andconcentrated under reduced pressure (bath temperature at 35° C.) toafford the crude 4,6-dichloropyrimidine-5-carbaldehyde (9, 270 g, 395 gtheoretical, 68.4%) as yellow-orange solid. A 20 g portion of this crudematerial was purified by Kugelrohr distillation (oven temperature at90-100° C., 225 mTorr) to give 15.3 g of pure4,6-dichloropyrimidine-5-carbaldehyde (9) as a white solid that turnedyellow on standing at room temperature. (On standing crude 9 undergoesslow hydrolysis with formation of HCl. Prior to use in the next stepcrude 9 was dissolved in a mixture of EtOAc and toluene and filtered toremove insoluble material. The filtrate washed with H₂O, saturatedNaHCO₃ solution, brine, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure and the resulting yellow solid used the followingday.) For 9: ¹H NMR (CDCl₃, 300 MHz) δ ppm 10.46 (s, 1H), 8.89 (s, 1H).

4-Amino-6-chloropyrimidine-5-carbaldehyde (10)

A solution of 7M NH₃ in MeOH (265 mL, 1.8602 mol, 2.0 equiv) was addedover 1.25 hr to a solution of 4,6-dichloropyrimidine-5-carbaldehyde (9,163.7 g, 0.9301 mol) in toluene (3 L). The reaction temperature slowlyincreased from 20 to 26° C. and a yellow suspension formed. Mild coolingwas applied to maintain the reaction temperature at ≦26° C. Thesuspension was stirred 3.5 hr at room temperature before the solids werecollected by filtration. The solids were washed with EtOAc (1 L). Thefiltrate was concentrated under reduced pressure, and the solids weretriturated with toluene/heptane (2:1 v/v, 600 mL), filtered and dried togive 71.1 g of 4-amino-6-chloropyrimidine-5-carbaldehyde (10) as ayellow solid. The original solid filtered from the reaction mixturecontained additional 10. The product was extracted from the filteredsolid by stirring in EtOAc (1.25 L) for 1.5 hr, filtering, then stirringin THF (750 mL) for 1 hr and filtering. Both EtOAc and THF filtrateswere concentrated under reduced pressure, and the resulting solids weretriturated with toluene/heptane (2:1 v/v, 450 mL), filtered and dried togive an additional 44.1 g of 4-amino-6-chloropyrimidine-5-carbaldehyde(10) as yellow solids. The combined yield of4-amino-6-chloropyrimidine-5-carbaldehyde (10, 115.2 g, 146.5 gtheoretical) was 78.6%. For 10: ¹H NMR (DMSO-d₆, 300 MHz) δ ppm 10.23(s, 1H), 8.71 (bs, 1H), 8.55 (bs, 1H), 8.39 (s, 1H); C₅H₄ClN₃O (MW,157.56), LCMS (EI) m/e 158 (M⁺+H).

6-Chloro-5-(2-methoxyvinyl)pyrimidin-4-ylamine (12)

A suspension of (methoxymethyl)triphenyl-phosphonium chloride (11, 276.0g, 0.807 mol, 1.1 equiv) in THF (1.5 L) was cooled in an ice/salt bathto −2° C. and 1M KO^(t)Bu in THF (807 mL, 0.807 mol, 1.1 equiv) wasadded over 1.5 hr at −2 to −3° C. The deep red-orange mixture wasstirred for 1 hr at −2 to −3° C.4-Amino-6-chloropyrimidine-5-carbaldehyde (10, 115.2 g, 0.7338 mol, 1.0equiv) was then added portion wise to the reaction mixture as a solidform using THF (200 mL) to rinse the container and funnel. During theaddition the reaction temperature increased from −3 to 13° C. and abrown color developed. When the reaction temperature dropped to 10° C.,the cooling bath was removed and the reaction mixture was allowed towarm to room temperature and stirred 42 hr. The reaction mixture wascooled to −2° C. before being quenched by the slow addition of saturatedNH₄Cl aqueous solution (750 mL). The mixture was concentrated underreduced pressure to remove most of the THF. The residue was partitionedbetween EtOAc (3 L) and H₂O (1 L). The organic phase was filtered toremove insoluble material at the interface, then extracted with 2N HCl(4×250 mL) followed by 3N HCl (2×250 mL). The combined HCl extracts wereback-extracted with EtOAc (500 mL) then filtered through Celite toremove insoluble material. The filtrate was cooled in an ice/brine bath,adjusted to pH 8 with a 6N aqueous NaOH solution and extracted withEtOAc (3×1 L). The combined EtOAc extracts were washed with brine (1 L),dried over Na₂SO₄, stirred with charcoal (10 g) and silica gel (10 g)for 1 hr. The mixture was filtered through Celite, washing the Celitepad with EtOAc (1 L). The filtrate was concentrated, co-evaporatingresidual EtOAc with heptane (500 mL). The resulting tan solid was pumpedunder high vacuum for 2 hr to afford crude6-chloro-5-(2-methoxyvinyl)pyrimidin-4-ylamine (12, 72.3 g, 136.2 gtheoretical, 53.1%). The crude 12 was used in the following reactionwithout further purification. A sample of crude 12 (2.3 g) was purifiedby chromatography on silica gel, eluting with 0-35% EtOAc/heptane togive 1.7 g of pure 12 as a white solid, which is a 1:2 mixture of E/Zisomers. For 12: ¹H NMR (DMSO-d₆, 300 MHz) for E-isomer: δ ppm 8.02 (s,1H), 7.08 (bs, 2H), 6.92 (d, 1H, J=13.1), 5.35 (d, 1H, J=13.0 Hz), 3.68(s, 3H) and for Z-isomer: δ ppm 8.06 (s, 1H), 7.08 (bs, 2H), 6.37 (d,1H, J=6.8 Hz), 5.02 (d, 1H, J=6.7 Hz), 3.69 (s, 3H); C₇H₈ClN₃O (MW,185.61), LCMS (EI) m/e 186/188 (M⁺+H).

4-Chloro-7H-[pyrrolo[2,3-d]pyrimidine (1)

Concentrated HCl (5 mL) was added to a solution of crude6-chloro-5-(2-methoxyvinyl)pyrimidin-4-ylamine (12, 70.0 g, 0.3784 mol)in THF (700 mL) and the resulting reaction mixture was heated to refluxfor 7.5 hr. On warming a light suspension was formed that graduallyre-dissolved. When the reaction was deemed complete, the reactionmixture was cooled to room temperature and stirred overnight. SolidNaHCO₃ (15 g) was added and the mixture was stirred for 1 hr at roomtemperature. Charcoal (7 g), silica gel (7 g) and Na₂SO₄ (20 g) wereadded and the mixture heated to 40° C. The mixture was cooled to roomtemperature and filtered through Celite, washing the Celite pad with THF(1 L). The filtrate was concentrated under reduced pressure and theresulting solid was dried under reduced pressure to afford crude4-chloro-7H-[pyrrolo[2,3-d]pyrimidine (1, 58.1 g, 58.1 g theoretical,100%) as a yellow-brown solid. This crude product was dissolved in EtOAc(1 L) at 50-55° C. and treated with activated charcoal (3 g). Themixture was filtered while warm through Celite, washing the Celite padwith warm EtOAc (250 mL). The filtrate was concentrated to about 500 mLand the suspension was allowed to stand overnight. The suspension wascooled to 0-5° C. for 2 h before the solids were collected byfiltration. The solids were dried to afford pure4-chloro-7H-[pyrrolo[2,3-d]pyrimidine (1, 54.5 g, 58.1 g theoretical,94%) as yellow-brown crystals. For 1: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm12.58 (bs, 1H), 8.58 (s, 1H), 7.69 (d, 1H, J=3.5 Hz), 6.59 (d, 1H, J=3.5Hz); LCMS (EI) m/e 154/156 (M⁺+H).

4-Iodopyrazole (14)

A flask equipped with a nitrogen inlet, addition funnel, thermowell, andmechanical stirrer was charged with pyrazole (13, 450 g, 6.62 mol) andtetrahydrofuran (5 L). The mixture was cooled to 10° C. andN-iodosuccinimide (NIS, 1490 g, 6.62 mol, 1.0 equiv) was added inportions as a solid. The reaction mixture (slight suspension) wasstirred at room temperature for 1 hour (longer reaction times may benecessary depending on ambient temperature). The mixture was thenfiltered and the THF was removed under reduced pressure. The residue wassuspended in ethyl acetate (6 L) and insoluble materials were filtered.The dark filtrate was sequentially washed with aqueous saturated sodiumthiosulfate solution (2×3 L) (organic layer lightens to a pale yellow),water (2×3 L), and brine (2 L). The organic layer was dried over sodiumsulfate, filtered, and concentrated under reduced pressure to afford4-iodopyrazole (14, 1138 g, 1284.1 g theoretical, 88.6%) as white topale yellow solids after being dried in a vacuum oven at 30° C.overnight. For 14: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 13.17 (bs, 1H), 7.93(bs, 1H), 7.55 (bs, 1H); C₃H₃₁N₂ (MW, 193.97), LCMS (EI) m/e 195 (M⁺+H).

1-Trimethylsilyl-4-iodopyrazole (15)

To a flask equipped with a reflux condenser, a nitrogen inlet,mechanical stirrer, and a thermowell was charged 4-iodopyrazole (14, 200g, 1.03 mol) and THF (2 L). To this solution was added triethylamine(TEA, 158 mL, 1.13 mol, 1.1 equiv) and the resulting solution was cooledto 0° C. in an ice-brine bath. To this solution was addedchlorotrimethylsilane (TMS-Cl, 137 mL, 1.08 mol, 1.05 equiv) with rapidstirring allowing the temperature to reach 18° C. (The reaction becomesvery thick and difficult to stir, but becomes manageable after overtime). When the exotherm had subsided, the cold bath was removed and thereaction was warmed to room temperature. The reaction was followed by GCand was found to be deemed complete after about 1 hour (Sampling ofreaction must be done out of air and diluted with dry solvent to preventTMS hydrolysis). The reaction mixture was then diluted with heptane (2L) before being filtered under nitrogen. The solvent was removed fromthe filtrate under reduced pressure venting the rotovap with nitrogen.The residual oil was diluted with heptane (1 L) and re-concentrated. Ifthe solids formed upon adding the heptane, a second filtration isnecessary. The residue was then distilled under the reduced pressure(70-90° C. at about 0.5 Torr) using a Kugelohr to afford1-trimethylsilyl-4-iodopyrazole (15, 263 g, 274.1 g theoretical, 96%) asa colorless oil. (This material must be kept under nitrogen at all timessince the TMS group rapidly hydrolyzes.) Subsequently, we have foundthat 1-trimethylsilyl-4-iodopyrazole can be prepared by heating theiodopyrazole (14) with 2 equivalents of hexamethyldisilazane for 1 hr.

4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (17)

A flask equipped with a mechanical stirrer, nitrogen inlet, additionfunnel and thermowell was charged with 1-trimethylsilyl-4-iodopyrazole(15, 225.1 g, 0.85 mol) and THF (2200 mL). This mixture was cooled to−6° C. in an ice/salt/brine bath and isopropyl magnesium chloride (2 Min THF, 510 ml, 1.02 mol, 1.2 equiv) was added at a rate such that thetemperature did not exceed 0° C. The extent of metal/halogen exchangewas monitored by GC and was found complete after about 10 min. To theorange brown solution was added2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(isopropylpinacolborate, 16, 347 mL, 1.7 mol, 2.0 equiv) slowly at firstkeeping the temperature below 0° C. and then fairly rapidly after about½ of the compound was added allowing the temperature to reach 5° C. (thereaction becomes quite thick and then thins out slowly). The reaction isthen stirred at 0° C. for 10 min before being warmed to room temperatureover 1 hr and stirred at room temperature for an additional 1 hr. Thereaction was cooled to 6° C. and saturated aqueous ammonium chloridesolution (2.2 L) was added with a temperature increase to 25° C. Themixture was stirred for 5 minutes before being diluted with toluene (10L). The layers were separated (a large amount of solid is present in theaqueous layer) and the organic layer was sequentially washed with water(6×2.2 L), brine (2×2.2 L), dried over sodium sulfate, filtered, andconcentrated under reduced pressure. Residual toluene was co-evaporatedwith heptane to afford4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (17, 90.3 g,164.9 g theoretical, 54.8%) as a white solid. For 17: ¹H NMR (DMSO-d₆,400 MHz) δ ppm 13.08 (bs, 1H), 7.94 (s, 1H), 7.62 (s, 1H), 1.23 (s,12H); C₉H₁₅BN₂O₂ (MW, 194.04), LCMS (EI) m/e 195 (M⁺+H).

1-(Ethoxyethyl)-4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(19)

A 22 L 4-neck flask equipped with a mechanical stirrer, thermowell,addition funnel, condenser and N₂ inlet was charged with4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (17, 1.42kg, 7.32 mol), toluene (9.5 L) and ethyl vinyl ether (18, 790.5 g, 1050mL, 10.98 mol, 1.50 equiv). A 4 M HCl in dioxane (50 mL) was added viaan addition funnel over 10 minutes and the resulting reaction mixturewas heated at 35-40° C. for 7 hr to give a clear homogeneous solution.When the reaction was shown to be complete by GC, solid NaHCO₃ (130 g)was added and the mixture was stirred for 1 hr before being filtered.The filtrate was concentrated under reduced pressure. Heptane (200 mL)was added to the residue to affect crystallization. The solid wascollected by filtration and dried in a vacuum oven to afford1-(ethoxyethyl)-4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(19, 1.896 Kg, 1.948 Kg theoretical, 97.3%) as a white to off-whitesolid. For 19: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.09 (s, 1H), 8.58 (s,1H), 7.62 (s, 1H), 5.55 (q, 1H, J=6.1 Hz), 3.37 (dq, 1H, J=7.1, 9.6 Hz),3.12 (dq, 1H, J=7.0, 9.7 Hz), 1.56 (d, 3H, J=6.0 Hz), 1.24 (s, 12H),1.00 (t, 3H, J=7.0 Hz); C₁₃H₂₃BN₂O₃ (MW, 266.14), LCMS (EI) m/e 267(M⁺+H).

1-(ethoxyethyl)-4-iodo-1H-pyrazole (20)

A 22 L 4-neck flask equipped with an mechanical stirrer, thermowell, N₂inlet and condenser was charged with 4-iodo-1H-pyrazole (14, 1.00 Kg,5.16 mol) and toluene (10 L) and ethyl vinyl ether (18, 557 g, 740 mL,7.73 mol, 1.5 equiv) was added. To the suspension 4 M HCl in dioxane (32mL, 0.128 mol, 0.025 equiv) was added over 5 min with formation of aslightly thicker white suspension. The mixture was heated carefully to35-40° C. at which point a mild exotherm to about 40° C. occurred withrapid dissolution of all solids to give a clear light yellow solution.The reaction mixture was heated at about 40° C. for an additional 0.5 hruntil the GC analysis indicated the reaction was complete. The solutionwas allowed to cool to 25-30° C. and solid NaHCO₃ (108 g, 1.29 mol, 0.25equiv) was added. The suspension was stirred for 1 hr at roomtemperature to ensure the complete neutralization of HCl. The mixturewas then filtered and the filtrate was concentrated under reducedpressure. The residual liquid was fractionally distilled to afford1-(ethoxyethyl)-4-iodo-1H-pyrazole (20, 1.346 Kg, 1.373 Kg theoretical,98%) as a pale yellow liquid (bp 89-93° at about 1 torr). For 20: ¹H NMR(CDCl₃, 250 MHz) δ ppm 7.61 (s, 1H), 7.47 (s, 1H), 5.46 (q, 1H, J=6.0Hz), 3.48-3.23 (m, 2H), 1.60 (d, 3H, J=6.0 Hz), 1.11 (t, 3H, J=7.0 Hz);C₇H₁₁IN₂O (MW, 266.08), LCMS (EI) m/e 267 (M⁺+H).

2-Isopropoxy-4,4,5,5-tetramethyl[1,3,2]dioxaborolane (16)

A 5 L 4-neck flask equipped with a reflux condenser, mechanical stirrer,N₂ inlet, and thermowell was flushed well with N₂ and charged withisopropyl borate (2.673 L, 11.5 mol, 1.15 equiv) and pinacol (1.179 kg,10 mol). The resulting mixture was heated at reflux (80-85°) forovernight. The mixture was then cooled to room temperature, transferredto a 5 L 4-neck flask equipped with a 24 inch Vigreux column, magneticstirrer, and thermowell. The mixture was distilled at atmosphericpressure under nitrogen. After the low boiling fraction (bp 90-180°)which contained predominately 2-propanol and isopropyl borate (GCanalysis) was removed, the completed distillation afforded2-isopropoxy-4,4,5,5-tetramethyl[1,3,2]dioxaborolane (10, 1.628 kg, 1.86Kg theoretical, 87.5%) as a colorless liquid (bp 180-185° C. with GCpurity >97.5%). This material was stored in Sure/Seal bottles tominimize hydrolysis.

1-(Ethoxyethyl)-4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(19)

A 22 L 4-neck flask equipped with a mechanical stirrer, thermowell,addition funnel, and N₂ inlet was charged with1-(ethoxyethyl)-4-iodo-1H-pyrazole (20, 700.0 g, 2.63 mol) and THF (5.5L). The resulting solution was cooled to between −12° C.-−15° C. Asolution of 2 M i-PrMgCl in THF (1513 mL, 3.03 mol, 1.15 equiv) wasadded via an addition funnel over 30 min while maintaining the reactiontemperature at <−5° C. and the tan suspension was stirred at <−5° C. for0.75 hr. The resulting reaction mixture was further cooled to −15° C.and 2-isopropoxy-4,4,5,5-tetramethyl[1,3,2]dioxaborolane (16, 734 g, 805mL, 3.95 mol, 1.5 equiv) was added rapidly via an addition funnel withthe reaction temperature increasing to ˜−5°. [Note: previous work withthe analogous TMS-protected pyrazole has shown that slow addition of2-isopropoxy-4,4,5,5-tetramethyl[1,3,2]dioxaborolane results in a loweryield.] A nearly clear light brown solution was developed followed byreformation of grayish light suspension. The cooling bath was thenremoved and the reaction mixture was allowed to warm to 16° C. over 0.75hr. The mixture was poured into 50 L reparatory funnel containing astirred saturated aqueous NH₄Cl solution (4 L). The mixture was dilutedwith toluene (8 L), heptane (8 L) and H₂O (2 L). The aqueous phase wasremoved and the organic phase was washed with warm (30° C.) H₂O (4×3 L)and saturated brine (2×3 L). The organic phase was dried over Na₂SO₄,and the solvents were removed under reduced pressure. The residualtoluene was further removed by co-evaporation with heptane (2 L). Theresidual oil was transferred to a 4 L beaker using a minimum amount ofheptane (100 mL) and scratched to induce crystallization. The solid wasfiltered, washed with heptane (200 mL) and dried overnight in a vacuumoven at 30-40° C. The filtrate was concentrated under reduced pressureand the residue was allowed to stand overnight. The resulting solid wasfiltered, washed with heptane (100 mL) and dried overnight in a vacuumoven at 30-40° C. The two crops were combined to afford1-(ethoxyethyl)-4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(19, 596 g, 700 g theoretical, 85.1%) as a white to off-white solid. For19: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.09 (s, 1H), 8.58 (s, 1H), 7.62 (s,1H), 5.55 (q, 1H, J=6.1 Hz), 3.37 (dq, 1H, J=7.1, 9.6 Hz), 3.12 (dq, 1H,J=7.0, 9.7 Hz), 1.56 (d, 3H, J=6.0 Hz), 1.24 (s, 12H), 1.00 (t, 3H,J=7.0 Hz); C₁₃H₂₃BN₂O₃ (MW, 266.14), LCMS (EI) m/e 267 (M⁺+H).

4-Bromopyrazole (21)

Pyrazole (13, 34.0 g, 0.5 mol) and NBS (89.0 g, 0.5 mol, 1.0 equiv) weresuspended in water (625 ml). The resulting suspension was stirred overnight at room temperature. The reaction mixture was then extracted withEtOAc (2×100 mL). The combined EtOAc extracts was washed with aqueousNa₂S₂O₃ and brine, dried over Na₂SO₄, and concentrated under reducedpressure to afford 4-bromopyrazole (21, 72.0 g, 73.5 g theoretical, 98%yield) as a white solid (GC purity: >98%).

4-Bromo-1-(ethoxyethyl)-1H-pyrazole (22)

To a solution of 4-bromopyrazole (21, 70.0 g, 0.476 mol) in CH₂Cl₂ (600mL) was added a solution of 3.1 M HCl in dioxane (4 mL) and ethyl vinylether (18, 41 g, 0.569 mol, 1.2 equiv). The resulting reaction mixturewas stirred at room temperature for 3 hrs. The reaction was quenchedwith aqueous NaHCO₃ and the two layers were separated. The organic layerwas washed with water, dried over Na₂SO₄, and concentrated under reducedpressure to dryness to afford 4-bromo-1-(ethoxyethyl)-1H-pyrazole (22,113 g, 104.3 g theoretical, 97% yield) as an oily (GC purity: 89%),which was directly used in the subsequent reaction without furtherpurification.

1-(Ethoxyethyl)-4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(19)

To a 100 ml solution of iPrMgCl.LiCl (50 mmol, 1.8 equiv) was added4-bromo-1-(ethoxyethyl)-1H-pyrazole (22, 6.15 g, 28 mmol) at roomtemperature. The resulting reaction mixture was stirred at roomtemperature for 12 hrs and then cooled to −20° C. Methoxy pinacolborate(23, 10.6 g, 67 mmol, 2.4 equiv) was then added to the reaction mixture.The resulting mixture was stirred at 0-10° C. for 1 h. Aqueous NH₄Cl wasadded to quench the reaction. The mixture was then extracted withpetroleum ether (PE). The combined PE extracts were washed withsaturated NaHCO₃, dried over Na₂SO₄ and concentrated under reducedpressure. The crude product was crystallized in PE to afford1-(ethoxyethyl)-4-(4,4,5,5-tetramethyl[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(19, 4.2 g, 7.45 g theoretical, 56.4% yield) as a white to off-whitesolid (GC purity: ˜99%). For 19: ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.09(s, 1H), 8.58 (s, 1H), 7.62 (s, 1H), 5.55 (q, 1H, J=6.1 Hz), 3.37 (dq,1H, J=7.1, 9.6 Hz), 3.12 (dq, 1H, J=7.0, 9.7 Hz), 1.56 (d, 3H, J=6.0Hz), 1.24 (s, 12H), 1.00 (t, 3H, J=7.0 Hz); C₁₃H₂₃BN₂O₃ (MW, 266.14),LCMS (EI) m/e 267 (M⁺+H).

A number of embodiments of the invention have been described.Nevertheless, it will be understood that various modifications may bemade without departing from the spirit and scope of the invention.Accordingly, other embodiments are within the scope of the followingclaims.

What is claimed is:
 1. A process of preparing a composition comprisingan enantiomeric excess of the (R)-enantiomer of a compound of FormulaIII′:

comprising: reacting said compound of Formula I″:

with boron trifluoride diethyl etherate, followed by aqueous ammoniumhydroxide to form a composition comprising an enantiomeric excess of the(R)-enantiomer of said compound of Formula III′; wherein * is a chiralcarbon.